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Full text of "Proceedings of the Indiana Academy of Science"

PROCEEDINGS 

of the 

Indiana Academy 
of Science 

Founded December 29, 1885 



Volume 90 
1980 



DONALD R. WINSLOW, Editor 

Indiana University 

Bloomington, Indiana 



Spring Meeting 
April 25, 26, 1980 

Geneva Center 
Rochester, Indiana 



Fall Meeting 

November 6, 7, 8, 1980 

Saint Joseph's College 

Rensselaer, Indiana 

Published at Indianapolis, Indiana 
1981 



1. The permanent address of the Academy is the Indiana Academy of Science, 140 North 
Senate Avenue, Indianapolis, Indiana 46204. 

2. Instructions for Contributors appear at the end of this volume. 

3. Exchanges. Items sent in exchange for the Proceedings and correspondence concern- 
ing exchange arrangements should be addressed: 

John Shepard Wright Memorial Library of the Indiana 

Academy of Science 

140 North Senate Avenue 

Indianapolis, Indiana 46204 

4. Proceedings may be purchased through the Library at $7.00 for each volume. 

5. Reprints of technical papers often can be secured from the authors. They cannot be supplied by the 
Library nor by the officers of the Academy. 

6. The Constitution and By-Laws reprinted from Vol. 74 are available to members upon application to 
the Secretary. Necrologies reprinted from the various volumes can be supplied to relatives and friends of 
deceased members by the Secretary. 

7. Officers whose names and addresses are not known to correspondents may be addressed care of the 
Library. 



Donald R. Winslow 

Chairman and Editor 
Rita Barr 
Charlotte Boener 
Ernest Campaigne 
Donald P. Franzmeier 
James Gammon 



Editorial Board 
1981 



James Kellar 
Benjamin Moulton 
John Pelton 
Bruce Renda 
Carl Sartain 
Bernard S. Wostman 



TABLE OF CONTENTS 

Page 

Officers and Committees for 1980 3 

Minutes of the Spring Meeting (Executive Committee) 16 

Minutes of the Spring Meeting (General Session) 18 

Minutes of the Fall Meeting (Executive Committee) 19 

Minutes of the Fall Meeting (General Session) 23 

Annual Financial Report 26 

Annual Report, Indiana Junior Academy of Science 30 

Biological Survey Committee Report 34 

Necrology 38 

New Members for 1980 41 

ADDRESSES AND CONTRIBUTED PAPERS 

Presidential Address 

"Scientific Research and Economic Indicators," 

Robert E. Henderson 45 

1980-81 "Speaker of the Year" Address 

"Progress in Resolving Food Safety Problems: Systematic Evaluation 
of GRAS Food Ingredients," 

Harry G. Day 63 

Anthropology 

Francis X. Grollig, S. J. — Details in Mexican Archaeology* 72 

C. Dean Higginbotham — The Indiana Packet: Ecotone and Cultural Boundary 
within the Lower Wabash Valley* 72 

Robert E. Pace, William Rheinhardt, and Daniel Thiel — Cataract Chert 

And Its Distribution In West-Central Indiana* 72 

Robert E. Pace — Stone Box Burials In West-Central Indiana* 72 

Curtis H. Tomak — A Note on the Distribution of Riverton Points* 72 

Curtis H. Tomak — Rock Shelters: An Important Archaeological Resource of 

Southern Indiana* 73 

Charles P. Warren — Two Case Studies in Forensic Anthropology: Calumet 

Township, Indiana; and Griffith, Indiana* 73 

James 0. Bellis — An Experiment With Time Control in Intensive Site 

Survey 74 

Paul Jean Provost — The Fate of the Soul in Modern Aztec Religious 

Thought 80 

Botany 

William W. Bloom -A Proposed Third Function for Root Caps* 86 

William Davies — Utilization of an Artificial Beach in Investigation of 
Phytopsammon communities* 86 

* Abstracts 



vi Table of Contents 

Page 

Stanley L. Burden, David Woodall, Kathleen Donica, Rick Thompson, 
Jon Condit, Steven Beeson, and Douglas Taylor — Laboratory Inter- 
facing with the Apple II Plus Microcomputer: A Computer Controlled 
Titrator with High Resolution Color Graphics Display* 174 

Marshall P. Cady, Jr. — Test of a Nonisothermal Apparatus Suitable for 
the Study of Energy and Mass Transport Coefficients* 175 

E. Campaigne and R. Yodice — The Schmidt Reaction of 3a,4,5,6-Tetrahydro- 
succinimido [3,4-b] acenaphthen-10-one and Its Alkylated Derivatives* .... 176 

R. J. Coffey, D. J. Reuland, and W. A. Trinler — The Identification of 
Synthetic Fibers of Forensic Interest by a Combination of Differential 
Thermal Analysis and Infrared Spectrophotometry* 176 

Joseph J. Gajewski and Kevin E. Gilbert — Energy Surfaces of Sigma- 
tropic Shifts* 176 

Robert H. L. Howe — The Chemistry of Cyanogen Compounds Degrad- 
ation* 176 

Mark D. McIntire, Paul L. Bock, John A. Mosbo, and Bruce N. Storhoff — 
Parameterization of the Empirical Molecular Conformation Approach 
CAMSEQ for Phosphines* 176 

Thomas W. Myers and Pang F. Ma — A Study of Adenosine Deaminase in 
Human Serum* 177 

Lois M. Ounapu, John A. Mosbo, Paul L. Bock, and Terry L. Kruger— 
Equilibria Between Diols and the NMR Shift Reagent Eu(fod) 3 * 177 

Shashi Puttaswamy and Pang F. Ma — A Comparative Study of the Inter- 
action Between the Conversion Factors from Human Tissues with the 
Small Forms of Adenosine Deaminase from Various Organisms* 177 

Brenda L. Schuffman and Eric R. Johnson — The Hydrolysis of Bovine 
Glucagon by a Denaturant-stable Protease* 178 

Eugene Schwartz, Mark Bradley, and Timothy Neufeld — Dielectric and 
Electronic Polarizations of Substituted Metal-Acetylacetone Complexes* . . 178 

Wanda J. Wills and Eric R. Johnson — The Isolation of a New Denaturant- 
stable Protease from a Commercial Protease Preparation, Pronase* 179 

Barth H. Ragatz — Interactions of Various Homopolypeptides with Human 
Platelet-Rich Plasma Suspensions 180 

James T. Streator, Pam Bertram, and Caryl L. Coy — Fluoride Content of 
Common Foods 186 

Ecology 

Robert L. Ball — Changes in Yellow Bass Growth Rates and Density During 

the First Ten Years of Its Establishment in Monroe Reservoir* 190 

Kenneth M. Brown — Habitat, Food, and Life History Overlap in Temporary 

Pond Snails: Evidence for Competition* 190 

Stephen R. Carpenter and Durland Fish — Detrital Dynamics Regulate 

Mosquito Production in Treehole Ecosystems* 191 

William Y. B. Chang — Limiting Nutrients and Primary Productivity* 191 

William Bliss Crankshaw — Attrition of White Ash in Red Pine Plantations 

in Eastern New York* 191 

• Abstracts 



Table of Contents v 

Page 
M. E. Jacobs — The Role of Beta-alanine in Pigmentation Polymorphisms* . . . jgp 

James S. Miller, Samuel 0. Ikharebha, Victor C. Gavino, George E. 
Milo, and David G. Corn well — Antioxidants, Fatty Acids and Oxidant 
Stress, and the Control of Cell Proliferation in Culture* 230 

Kenneth C. Miller and Stanley N. Grove — Effects of Cytochalasins on 
Selected Species of Aspergillus and Mucor* y^ 

Dorothy M. Morre, D. James Morre, and Martha Walter — Effect of 
Vitamin A on the Hepatic Golgi Apparatus Architecture* 13] 

Dorothy M. Morre and Ann L. Rosenthal — Distribution of Acid Phos- 
phatase in Transplanted Hepatomas of the Rat* 232 

D. James Morre, Edward M. Croze, Cheryl Franz, and C. M. Eppler — 
Exocytosis: Routes and Kinetics of Delivery of Secretory and Membrane 
Proteins* 132 

Michael J. Stewart and Charles W. Goff — Comparative Cytochemical 
Studies of Onion Root Tip with Specific and Non-specific Substrates for 
Nucleoside Diphosphatase(s)* J32 

James A. Sweigard and Stanley N. Grove — Cytochalasin A (0.1-1 ug/ml) 
rapidly halts Hyphal Tip Growth in Rhizoctonia solani* 233 

Martha Twaddle, Ralph A. Jersild, and D. James Morre — Kinetics of 
Appearance of Lipoprotein Particles in Cisternae of Subsurface Smooth 
Endoplasmic Reticulum of Isolated Rat Livers Perfused with Free Fatty 

Acid * 133 

Martin A. Vaughan and J. P. Braselton — Persistent Nucleoli in Various 
Meristems of Phaseolus aureus (mung bean)* 234 

Henry C. Womack and Mohinder S. Jarial — The Effect of Sympathectomy 
on the Structure of the Rat Pineal Gland* 234 

Vernon G. Archer and James P. Holland — Effect of Maternal Thyroid 
Activity Upon In Vitro Protein Synthesis in the Rat Blastocyst 236 

S. D. Barnard, S. D. Warner, W. J. Caldwell, and F. Parenti — Chemi- 
cally Induced Hepatocellular Inclusions Associated with Tumorigenicity 
in the Liver of the Rat 243 

Theodore A. Craig and Frederick L. Crane — Evidence for A Trans-Plasma 
Membrane Electron Transport System in Plant Cells 250 

D. K. Hansen and M. E. Hodes — Teratogenic Potential of Phenytoin in 
Different Strains of Mice 256 

Gary R. Matyas, Dorothy A. Werderitsh, Dorothy M. Morre, and D. 
James Morre — Comparison of Metastatic and Non-metastatic Transplant- 
able Tumors from the Jaw Region of the Rat 2(31 

Chemistry 

Randall K. Atkins, Paul L. Bock, John A. Mosbo, and Bruce N. Stor- 
hoff — Comparisons of the Semi-empirical Molecular Orbital Treatments 
MINDO/3 and MINDO with Some Simple Phosphines* 174 

John E. Bartmess, Robert L. Hays, and Stephen R. Wilson — Gas Phase 
Chemistry of 1,3-Dithiane* 174 



Abstracts 



iv Table of Contents 

Page 

Gina Fernandez, Jay H. Jones, and David L. Dilcher — A Developmental 
Study of Foliar Epidermal Features in Castanea mollissima Blume 
(Fagaceae)* 86 

Stephen W. Fletcher — Effects of Irradiance on the Morphological Charac- 
teristics of Two Plant Species of the Maritime Strand* 87 

Vonda Frantz and Kathryn Wilson — The Ultrastructure of Nonarticulated 
Branched Laticifers in Asclepias tuberosa L. (Butterfly Weed)* 87 

Jay H. Jones and David L. Dilcher — Paleobotanical Nomenclature: Prin- 
ciples, Problems and Proposals* 88 

Stephen R. Manchester and David L. Dilcher — Fruits of the Pterocarya 
alliance (Juglandaceae) from the Paleogene of the Rocky Mountain 
Region* 88 

John L. Roth, Jr., David L. Dilcher, and Brent A. McKim — Stomatal 
Development in Asimina triloba (L.) Dunal* 89 

Robert Schwarzwalder, Jr. and David L. Dilcher — Bisexuality in the 
Pistilate Inflorescence of Platanus occidentalis L.* 89 

James P. Shepard and William R. Chaney — Endomycorrhizae Increases 
Growth of Sycamore Seedlings* 90 

Marilyn Sue Veselack and Jerry J. Nisbet — The Distribution and Uses of 
Arundo donax* 90 

Robert D. Williams — Squirrel Resistant Walnuts for Direct Seeding? Not 
Yet* 90 

Wayne T. Williams and Roland W. Usher — Air Toxicity to Eastern White 
Pines in Indiana* 91 

Rita Barr and Frederick L. Crane — Phosphate Stimulates or Inhibits Sili- 
comolybdate Reduction in Spinach Chloroplasts 92 

Deborah Williams Craton and Robert D. Williams — Juglone Dermatitis: 
Allergy or Irritant? 98 

Gary E. Dolph, Jon Bolke, and Jill Rodaer- Woody Vegetation of the 
Cass County Outdoor Education and Wildlife Area 103 

Gail E. Evans-Ruhl Donald H. Scott, and Paul C. Pecknold-A Compila- 
tion of Plant Diseases and Disorders in Indiana — 1980 107 

Heidi A. Schmitt and Connie M. Weaver — Chromium-Zinc Interaction in 
Accumulation of Minerals by Bush Beans 125 

Cell Biology 

John H. Elder and D. James Morre — Observations on the Tertiary Struc- 
ture of Ribosomes Released from Membranes of Rough Endoplasmic Reti- 
culum* 129 

David C. Evers and D. James Morre — Complex Gangliosides Bind Fib- 
ronectin* 129 

D. K. Hansen, W. Chaekal, and M. E. Hodes — Electrophoretic Study of the 
Deoxyribonuclease Activity of Human Urine* 129 

Zafar Iqbal — Characterization of A Fast Transported Calcium-Binding Pro- 
tein in Mammalian Nerve* 130 



Abstracts 



Table of Contents vii 

Page 
William Bliss Crankshaw — Success of Bald Cypress Seedlings in the 

Drawdown Zone at Salamonie Reservoir* 191 

Dennis Devries — Effect of Habitat Productivity, Permanence, and Preda- 

tion on the Life History of a Temporary Pond Snail* 192 

John S. Fezy and Richard W. Greene — Periphy ton Productivity of Three 

Sample Sites along Juday Creek, St. Joseph County, Indiana* 192 

Stephen W. Fletcher — An Updated Evaluation of Sampling Efficiencies of 

Overstory Sampling Methods* 192 

Paul A. Glander and Thomas S. McComish — Macrophyte Induced Fluctua- 
tions of Water Chemistry in and East-Central Indiana Borrow Pit Lake* . . 193 
Joanne M. Payton and Richard W. Greene — A Comparison of the Effect of 

Aluminum on a Single Species Algal Assay and Indigenous Community 

Algal Toxicity Bioassay* 193 

Ken Roberts and W. Herbert Senft II — Growth and Phosphorus Uptake 

as A Function of Temperature in the Colonial Green Alga Volvox globator 

L* 194 

Robert K. Rose — The Small Mammals of Spencer County, Indiana* 194 

Robert Schwarzwalder, Jr. — Trends in the Climatic Adaptations of Lichens* 194 
Bruce W. Schwenneker and Ronald A. Hellenthal — Seasonal, Spatial and 

Developmental Variability of Benthic Macroinvertebrates in A Northern 

Indiana Stream* 195 

W. Herbert Senft II and Arthur J. White — Spatial Patterning of A Volvox 

globator L. Population in A Northern Minnesota Pond* 195 

Patrick F. Sullivan and Stephen R. Carpenter — Relationships of Algal 

Trophic State Indices in Indiana Lakes and Reservoirs* 196 

Forrest C. Brown and Thomas S. McComish — Fishes Reported from Waters 

of the Hoosier National Forest 197 

Clarence F.Dineen — Fishes of Spicer Lake 204 

William L. Fisher and James R. Gammon — The Implications of Rotenone 

Eradication on the Fish Community of Eagle Creek in Central Indiana 208 

C. L. Rhykerd, E. L. Hood, B. 0. Blair, Ekpo Ossom, and J. W. Lightner — 

Alopecurus pratensis L. Discovered in Porter County 216 

Engineering 

Warren W. Bowden — Development of a Robust Routine for Calculating 

Vapor-Liquid Equilibrium Conditions* 219 

Christopher B. Burke -Combined Sewer Separation: Case Study* 219 

David D. Chesak — Microcomputer Driven Multi-Point Controller* 220 

Wayne F. Echelberger, Jr., J. Michael Jeter, Steve W. Kim, and J. C. 
Randolph — Chemical and Physical Characterization of Wastewater 

Sludges in Indiana* 220 

Aldo Giorgini and Mark Smith -Particulate Sedimentation in Shallow 
Lakes* 221 



Abstracts 



viii Table of Contents 

Page 
Robert H. L. Howe — The Boundary Conditions of Oxygen Transfer in Water 

with Respect to Temperature* 221 

Robert H. L. Howe and Hakki Dingil — The Determination of the Rising 

Velocity of a Gas Bubble Through a Liquid Column by Howe's Method* ... 221 
P.G. Katz and G. H. Tobes - Difference Models of River Reaches.* 222 

A. Ramachandra Rao — Analysis of Suspended Particulate Data from Chi- 
cago* 222 

Gerard F. Sheldon and Donald D. Gray — Performance of the Purdue 
Hydromechanics Laboratory Closed Circuit Wind Tunnel* 222 

Bithin Datta and Mark H. Houck — Grain Reserve Systems: Failure of 
Multiobjective Decisions Under Catastrophic Intervention, A Case for 
Topologic Stability in Singular Mapping 224 

Mohammad Karamouz and Mark H. Houck — Reservoir Operating Rules . . . 230 

Entomology 

William E. Chaney, Donald L. Schuder, and David K. Reed — Notes on the 
Biology and Oviposition of the Twobanded Japanese Weevil (Callirhopalus 
bifasciatus Roelofs)* 234 

A. Yumi Clemons and Raymond J. Russo — Laboratory and Field Evaluation 
of Oviposition Preference in Toxorhynchites rutilus rutilus* 234 

Rosemarie Funkhouser and Raymond J. Russo — Clinical Variation of Tem- 
perature Dependent Survivorship in the Mosquito Aedes sollicitans* 234 

John W. Hart — Spring-tailed Insects of the Genus Proisotoma, subgenus 
Appendisotoma, from Manlove Woods* 235 

Sue Henderson and Robert R. Pinger — Oviposition and Larval Develop- 
ment of Toxorhynchites brevipalpis (Diptera: Culicidae) at Selected pH 
Levels* 235 

Bridget Hoban and George B. Craig, Jr. — Effectiveness of the Ovitrap for 
Monitoring Culex Populations* 235 

John Keltner and W. P. McCafferty — The Burrowing Mayflies (Ephemer- 
oidea) of Indiana* 236 

Stephen R. Madigosky, Robert R. Pinger, and Horst F. Siewert — Effects 
of pH on Oviposition Preference and Larval Development of Mosquito 
Species Aedes triseriatus* 236 

Raymond Russo and Kimberly Raubenheimer — Effect of Prey Density on 
the Use of Toxorhynchites rutilus as A Biological Control Agent* 237 

Alan C. York and Ronald D. Gardner — Home Garden Insecticide Use in 
Indiana* 237 

Stephen L. Durso and George B. Craig, Jr. — Treehole Productivity for 
Aedes triseriatus in Northern Indiana, 1975 vs. 1979 238 

Lorraine Leiser — Distribution of Aedes triseriatus (Say) in an Urban Area: 
Comparison of Two Survey Methods 248 

Philip T. Marshall and John J. Fa vinger — Indiana Pine Wilt Nematode 
Survey 254 



* Abstracts 






Table of Contents ix 

Page 
Robert W. Meyer — Insects and Other Arthropods of Economic Importance 

in Indiana During 1980 259 

B. Elwood Montgomery — Effect of Photoperiod and Temperature upon the 

Growth Rate of the Later Instars of Erythemis simplicicollis (Say) 

(Odonata; Libellulidae) 266 

David B. Taylor — New Distribution Records for Mosquitoes (Diptera: 

Culicidae) in St. Joseph County, Indiana 274 

Environmental Quality 

Thad J. Godish — Acid Precipitation Control Strategies* 281 

Thad J. Godish — Sensitivity of Weed Species to Ozone Exposures* 281 

Thad J. Godish, Virgil Konopinski, and David Peterson — Formaldehyde 
Air Pollution in Residential Housing* 281 

Robert H. L. Howe — The Chemical Properties of Biological Sludges* 282 

Robert H. L. Howe -The Meaning of pH in Environmental Control and 
Wastes Treatment* 282 

Patrick Sullivan -A Groundwater Quality Study of the Franklin County 
Sanitary Landfill, Brookville, Indiana* 282 

Roland W. Usher and Edmund A. ScHOFiELD-The Effects of Acid Mists on 
Nitrogen-Fixing Blue-Green Algae* 282 

Thad J. Godish — Effects of Nitric Oxide, Nitrogen Dioxide and Nitric Oxide- 
Nitrogen Dioxide Pretreatments on the Sensitivity of Tomato cv. Rutgers 
to Ozone 283 

D. W. Nelson and E. D. Orme — Phosphorus Status of Selected Indiana Lake 
and Reservoir Sediments 287 

Joseph R. Siefker and David P. McCleary — Chemical Analysis of Water 
Samples from Three Lakes in the Greene-Sullivan State Forest and the 
Wabash River at Clinton, Indiana 293 

Geology and Geography 

Konrad J. Banaszak — The pH of Precipitation in Indianapolis* 296 

Thomas J. Gallagher, Margaret M. Kimmel, and Gary W. Barrett — A 

Social Impact Assessment of the Brookville Reservoir on Union County, 

Indiana* 296 

Gerald E. Greengold and Terry R. West — Engineering Geology of an 

Operating Area Strip Mine, Southwestern Indiana* 297 

Joseph G. Hailer — Groundwater Chemistry in Vigo County, Indiana* 297 

Eric Koglin and Donald W. Ash — The Sedimentation of Lumsden Pond, 

Vigo County, Indiana* 298 

John B. Patton and Richard K. Leininger — Outlook for Shale-Oil Recovery 

from New Albany Strata in Indiana* 298 

Thomas Frank Barton — Rurbanization and the Countryside Urban Web in 

Indiana 299 

S. N. Ghose — A Statistical Correlation of Sulfur Content in Coal 5 and the 

Overlying Gray Shale 306 

* Abstracts 



x Table of Contents 

Page 
Andrea Sjoreen and Richard L. Powell -An Analysis of Irregularity of 

Surveyed Sections in Indiana 313 

Dan M. Sullivan, Curtis H. Ault, and George F. TANNER-Faulting in 

Perry and Spencer Counties, Indiana 323 

History of Science 

R. William Orr- Charles Lyell's Geologic Observations in Indiana, 1846* . . . 329 

Gene Kritsky — Mendel and the Origin of Species 330 

Walter H. Pierce -Arthur J. Phinney, M.D., Indiana's First Subsurface 
Geologist 335 

Microbiology and Molecular Biology 

William W. Baldwin, Ming Tseng, and Marshall Landay — A Simplified 
Method for Storing Anaerobic Bacteria* 340 

Kenneth F. Bartizal, Margaret H. Beaver, and Bernard S. Wostmann — 
The Effects of a Hexaflora on the Morphology of the Gerbil* 340 

Larry Herrman and Mary Lee Richeson — Isolation, Identification, and 
Reinfection of Cephalosporium spp. and Two Bacteria Associated with 
Wilt in Helianthus annuus* 341 

Robert H. L. Howe — Dissolved Oxygen Profile of an Aerobic Bio-Reactor* . . 341 

Mark 0. Oster and Sylvia Brehm Oster — Search for Phosphoproteins in 
Bacillus subtilis* 341 

Toni L. Poole and Carl Warnes — Rate Studies on Microbial Chitin Decom- 
position in the Freshwater Habitat* 342 

Mary Lee Richeson and Larry Herrman — Isolation and Identification of 
Two Bacteria Associated with Wilt in Helianthus annuus* 342 

Dilipkumar Vyas and Bruce Storhoff — Methodolgy for Measuring Nitro- 
gen Fixation by Acetylene Reduction in Beijerinckia and Klebsiella in 
Stream Litter Decomposition* 343 

Dilipkumar Vyas, Donald Hendrickson, Carl Warnes, and Bruce Stor- 
hoff— Investigation of the Role of Beijerinckia and Klebsiella as Nitro- 
gen Fixers in Stream Litter Decomposition* 343 

Donald Hendrickson — Bacterial Community Dynamics and Species Diver- 
sity of Slime Producing Bacteria at Selected Sites on the Ohio River 344 

Donald Hendrickson and Thomas S. McComish — Analysis of Factors 
Influencing Population Dynamics of Slime Forming Bacteria at Selected 
Sites on the Ohio River 351 

I. L. Sun and D. P. Gustafson — Properties of Defective Interfering Particles 
Induced by Photodynamic Treatment on Pseudorabies Virus 357 

Physics and Astronomy 

L. Dwight Farringer — A Valveless Drain-Down Solar Collector System* . . . 355 
J. W. Kaellner and D. R. Ober — Design and Construction of a Dye Laser* . 355 
Ronald E. Pitts — Some Problems Involved in Obtaining Accurate Oscillator 
Strengths for Use in Astrophysics* 366 



• Abstracts 



Table of Contents xi 

Page 
Roger L. Scott — IUE Observations of the Peculiar Object BL Lacertae* . . . 33$ 
R. Taylor and R. Cosby — Active Solar Heating Systems for Residential 

Application in Indiana: A Comparative Optimization Study* 3(57 

Robert L. Tonkel and Roger L. Scott- A Tube-Type 

Photographic Sensitometer* 367 

William W. Davis -"Contra Luz Opal:" Structure and 

Optical Properties 368 

Gerald P. Thomas and Mark Coggins- Microcomputer 

Assisted Coulomb Scattering Measurements in Nuclear 

Emulsion Pellicles 375 

Plant Taxonomy 

Buddy Cantrell — The Flora of the Southeastern United States: A Re- 
view* 382 

Theodore J. Crovello and Clifton Keller — An Updated, Computer-based 

Checklist of the Vascular Plants of Indiana: The Unending Synthesis* .... 382 

Richard J. Jensen — Phylogenetic Reconstruction in Quercus* 383 

Victor L. Riemenschneider — Vascular Plants of Sand Hill Nature Preserve, 

Pulaski County, Indiana* 383 

John L. Roth, Jr. and David L. Dilcher — The Epidermal Anatomy of Deer- 

ingothamnus Small and Its Taxonomic Implications* 384 

Helene Starcs— Najas marina L. in Indiana* 384 

John A. Bacone, Cloyce L. Hedge, and Theodore J. Crovello — The Status 

of Indiana's Rarest Plants: A Revision of the List of Endangered and 

Threatened Vascular Plants 385 

Mary Patricia Coons — Plant Distribution Records for Rush, Shelby and 

Decatur Counties 388 

John E. Ebinger and John A. Bacone — Vegetation Survey of Hillside Seeps 

at Turkey Run State Park 390 

William E. McClain — Additions to the Flora of Gibson County, Indiana .... 395 
William E. McClain — Additions to the Flora of Vermillion County, Indiana . 398 

Science Education 

Austin E. Brooks and William N. Doemel — Wabash Summer Aquatic 
Biology Program — The First Year* 403 

Richard A. Davis — Botanical and Zoological Prints in the Collections of the 
Hugh Thomas Miller Rare Book Room, Irwin Library, Butler University* . 403 

James D. Haddock — A Simple Quantitative Method for Teaching Phylo- 
genetic Relationships Among Animal Phyla to College Freshmen* 403 

John A. Ricketts and Paul B. Kissinger - STEP: The First Year* 404 

Stanley S. Shimer — Summary Report of Science Educators' Survey of 
Science Methods Curriculum* 405 

Patricia Arnett Zeck — The Necessity and Methods of Teaching Communi- 
cation Skills for Science Fair Participants* 405 



xii Table of Contents 

Page 
Soil and Atmospheric Sciences 

E. A. Matney and R. V. Ruhe — Clay Mineral Study of Soils from the Savan- 
nah River Plant South Carolina* 406 

Allan K. Nickell and Stanley M. Totten — Parent Material — Landscape — 
Soil Interrelationships in Jefferson County, Indiana* 406 

Lawrence A. Schaal — The Varying Length of the Growing Season in 
Indiana* 407 

Robert F. Dale, Beth C. Reinke, and John E. Wright — Freeze-Thaw Cycles 
in Indiana Soils 408 

J. J. King and D. P. Franzmeier — Morphology, Hydrology, and Management 
of Clermont Soils 416 

J. W. Lightner, C. L. Rhykerd, E. L. Hood, G. E. Van Scoyoc, and D. B. Men- 
gel— Effect of NPK Fertilization on Dry Matter Yield and Crude Protein 
Concentration of Poa pratensis L. Grown on Muck Soil 423 

G. C. Steinhardt, D. P. Franzmeier, and S. C. Valentine — Effect of Hydro- 
gen Peroxide Pretreatment on Particle-Size Analysis 428 

Russell K. Stivers — Fraction Sizes Change Available Phosphorus and Potas- 
sium Soil Test Values 435 

Zoology 

Larry L. Ganion — A Freeze-Fracture Study of the Mouse Ovarian Follicle and 
Zona Pellucida* 439 

John D. Goodman — The Northern Ringneck Snake A Host of Cepedietta 
sp.* 439 

G. R. Hogan — Variation of Lead Acetate-Induced Mortality in Estradiol- 
Treated Male Mice* 439 

Mohinder S. Jarial — The Fine Structure of the Rectal Pads of Grylloblatta 
compodeiformis* 440 

Doyal R. Lank, Jr. and Alice S. Bennett — Fatty Acid Distribution in Sala- 
manders of the Family Plethondontidae* 441 

Richard 0. McCracken, Peggy A. Johnson, and Doris D. Taylor— Biochemi- 
cal Effects of Tioxidazole on Hymenolepis diminuta in vivo* 441 

Ronald L. Richards — Vertebrate Remains from Carcass Crypt Cave, 
Lawrence County, Indiana* 442 

Henry Tamar — The Movements of A Jumping Rotifer, Polyarthra sp.* 442 

Barbara A. Brown — Extra Limbs in the Small-Mouthed Salamander, Amby- 
stoma texanum 443 

Thomas R. Kozel, Gordon K. Weddle, Kathy L. Welborn, John T. Dailey, 
Don C. Dailey, and Melvin W. Denner — A Fish Faunal Survey of Posey 
County, Indiana 446 

Ginger Lee Marenchin and David M. Sever — Survey of the Fishes of the 
St. Joseph River Drainage in St. Joseph and Elkhart Counties, Indiana .... 454 

John 0. Whitaker, Jr., Wynn W. Cudmore, and Barbara A. Brown — Foods 
of Larval, Subadult and Adult Smallmouth Salamanders, Ambystoma 
texanum, from Vigo County, Indiana 461 

* Abstracts 



Table of Contents xiii 

Page 

Instructions for Contributors 465 

Index 467 



Proceedings 

of the 

Indiana Academy 

of Science 



INDIANA ACADEMY OF SCIENCE 
Officers and Committees 1980 



OFFICERS 



President Robert E. Henderson, Indianapolis Center for Advanced 

(1980) Research 

1219 West Michigan Street, Indianapolis, Indiana 46202 

President-Elect Ralph A. Llewellyn, Department of Physics 

(1980) Indiana State University, Terre Haute, Indiana 47809 

Secretary John H. Meiser, Department of Chemistry 

(1980-82) Ball State University, Muncie, Indiana 47306 

Treasurer John A. Ricketts, Department of Chemistry 

(1979-81) DePauw University, Greencastle, Indiana 46135 

Director of Public 

Relations Walter A. Cory, Jr., Coordinator for School Science 

(1979-81) Memorial Hall West 108, Indiana University, 

Bloomington, Indiana 47405 

Editor Benjamin Moulton, Department of Geography and Geology 

(1978-80) Indiana State University, Terre Haute, Indiana 47809 



SECTION CHAIRMEN AND CHAIRMEN-ELECT FOR 1980: 

Anthropology 

Chairman Francis X. Grollig, Department of Anthropology 

Loyola University, Chicago, Illinois 60626 

Chairman-Elect Charles P. Warren, Department of Anthropology 

University of Illinois at Chicago Circle, 
Chicago, Illinois 60680 

Botany 

Chairman Gary Dolph, Department of Botany 

Indiana University at Kokomo, Kokomo, Indiana 46901 

Chairman-Elect Charles T. Hammond, Department of Botany 

St. Meinrad College, St. Meinrad, Indiana 47577 

Cell Biology 

Chairman . .Stanley N. Grove, Department of Biology 

Goshen College, Goshen, Indiana 46526 

Chairman-Elect Kara W. Eberly, Department of Biology 

St. Mary's College, Notre Dame, Indiana 46556 

3 



4 Indiana Academy of Science 

Chemistry 

Chairman Edward Miller, Department of Chemistry 

Manchester College, North Manchester, Indiana 46962 

Chairman-Elect Donald G. Clemens, Department of Chemistry 

Goshen College, Goshen, Indiana 46526 

Ecology 

Chairman W. Herbert Senft, Department of Biology 

Ball State University, Muncie, Indiana 47306 

Chairman-Elect Richard W. Greene, Department of Biology 

University of Notre Dame, Notre Dame, Indiana 46556 

Engineering 

Chairman Donald D. Gray, School of Civil Engineering 

Purdue University, West Lafayette, Indiana 47907 

Chairman-Elect Mark H. Houck, School of Civil Engineering 

Purdue University, West Lafayette, Indiana 47907 

Entomology 

Chairman Michael Sinsko, Indiana State Board of Health 

1330 West Michigan Street, Indianapolis, Indiana 46206 

Chairman-Elect David K. Reed, Fruit and Vegetable Insect Research 

Laboratory 
USDA-ARS Box 944, Vincennes, Indiana 47591 

Environmental Quality 

Chairman Howard Dunn, Department of Chemistry 

Indiana State University at Evansville, Evansville, 

Indiana 47712 

Chairman-Elect David Peterson, Indiana State Board of Health 

1330 West Michigan Street, Indianapolis, Indiana 46206 

Geology and Geography 

Chairman Kenneth R. Brehob, Department of Earth Sciences 

University of Notre Dame, Notre Dame, Indiana 46556 

Chairman-Elect Henry H. Gray, State Geological Survey 

611 North Walnut Grove, Bloomington, Indiana 47405 

History of Science 

Chairman Everett F. Morris, Dean, Academic Advisors 

Louisiana State University, Alexandria, Louisiana 71304 

Chairman-Elect Gene Kritsky, Department of Biology 

Tri-State University, Angola, Indiana 46703 

Microbiology and Molecular Biology 

Chairman Donald A. Hendrickson, Department of Biology 

Ball State University, Muncie, Indiana 47306 

Chairman-Elect Dorothy Adalis, Department of Biology 

Ball State University, Muncie, Indiana 47306 



Officers and Committees 5 

Physics and Astronomy 

Chairman Gerald P. Thomas, Department of Physics and Astronomy 

Ball State University, Muncie, Indiana 47306 

Chairman-Elect L. Dwight Farringer, 1407 East Orchard Drive 

North Manchester, Indiana 46962 

Plant Taxonomy 

Chairman John A. Bacone, Director, Division of Nature Preserves 

Indiana Department of Natural Resources 
601 State Office Building, Indianapolis, Indiana 46204 

Chairman-Elect . Robert J. Jensen, Department of Biology 

St. Mary's College, Notre Dame, Indiana 46556 

Science Education 

Chairman William G. Wert, Department of Life Sciences 

Indiana State University, Terre Haute, Indiana 47809 

Chairman-Elect Charles Gehring, Department of Life Sciences 

Indiana State University, Terre Haute, Indiana 47809 

Soil and Atmospheric Sciences 

Chairman Donald P. Franzmeier, Department of Agronomy 

Purdue University, West Lafayette, Indiana 47907 

Chairman-Elect Robert F. Dale, Department of Agronomy 

Purdue University, West Lafayette, Indiana 47907 

Zoology 

Chairman Robert R. Pinger, Department of Physiology and 

Health Sciences 
Ball State University, Muncie, Indiana 47306 

Chairman-Elect Larry R. Ganion, Department of Physiology and 

Health Sciences 
Ball State University, Muncie, Indiana 47306 

EXECUTIVE COMMITTEE, COUNCIL AND BUDGET COMMITTEE 



John A. Bacone 
*Otto K. Behrens 

William Beranek, Jr. 

Kenneth R. Brehob 

Lois Burton 

Ernest E. Campaigne 

Richard L. Conklin 
*Donald J. Cook 

Robert H. Cooper 

Walter A. Cory, Jr. 

Theodore Crovello 

Fay K. Daily 
♦William A. Daily 
*Harry G. Day 



Executive Committee 

♦Clarence F. Dineen 
William N. Doemel 
Gary Dolph 
Howard Dunn 
William R. Eberly 
Donald P. Franzmeier 

♦Raymond E. Girton 
Donald D. Gray 
Francis X. Grollig 
Stanley N. Grove 

♦Arthur T. Guard 

♦Frank A. Guthrie 
Robert E. Henderson 
Donald A. Hendrickson 



Earl A. Holmes 
♦William B. Hopp 

Marion T. Jackson 
♦Willis H. Johnson 

Duvall A. Jones 

Barry C. Knisley 
♦Alton A. Lindsey 

Ralph A. Llewellyn 
♦C. A. Markle 

Andrew G. Mehall 

John H. Meiser 
♦Melvin G. Mellon 
♦A. H. Meyer 
♦Howard H. Michaud 



6 



Indiana Academy of Science 



Edward Miller 
Benjamin Moulton 

♦Jerry J. Nisbet 
Robert R. Pinger 

♦John B. Patton 

♦S. N. Postlethwaite 

♦H. M. Powell 



John A. Ricketts 
Philip A. St. John 

♦DAMIAN V. SCHMELZ 

W. Herbert Senft 
Michael Sinsko 
Gerald P. Thomas 
'Past President of Academy 



♦W.J.Wayne 
♦J. Dan Webster 
♦Winona H. Welch 
William G. Wert 
Donald R. Winslow 
♦Howard R. Youse 



Council 

President — Robert E. Henderson 

President-Elect — Ralph A. Llewellyn 

Secretary- John H. Meiser 

Treasurer — John A. Ricketts 

Director of Public Relations — Walter A. Cory, Jr. 

Editor — Benjamin Moulton 

Chairman, Science & Society Committee- William Beranek, Jr. 

Budget Committee 

President -Robert E. Henderson 

President-Elect -Ralph A. Llewellyn 

Immediate Past President — J. Dan Webster 

Secretary — John H. Meiser 

Treasurer — John A. Ricketts 

Director of Public Relations — Walter A. Cory, Jr. 

Editor — Benjamin Moulton 

Chairman, Science & Society Committee — William Beranek, Jr. 

Co-Director, Junior Academy of Science — Keith Hunnings 

Co-Director, Junior Academy of Science — Leota Skirvin 

Co-Chairman, Program Committee — Duvall A. Jones 

Co-Chairman, Program Committee — Andrew G. Mehall 

Chairman, Library Committee — Lois Burton 

Chairman, Youth Activities Committee — Donald R. Winslow 

Chairman, Academy Foundation — William A. Daily 

Chairman, Research Grants Committee — Ernest E. Campaigne 



ELECTED COMMITTEES 

Academy Foundation 

William A. Daily, Chairman 5884 Compton Street 

(1979-80) Indianapolis, Indiana 46220 

Clyde R. Metz Department of Chemistry 

(1980-81) Indiana University-Purdue University 

1201 East 38th Street 
Indianapolis, Indiana 46205 

Bonding Committee 

Barry C. Knisley, Chairman 6022 Finlandia Lane 

(1980) Mechanicsville, Virginia 23111 

Donald R. Winslow Department of Science and Environ- 

(1980) mental Education 

Indiana University 
Bloomington, Indiana 47405 



Officers and Committees 7 

Research Grants Committee'" 

Ernest E. Campaigne, Chairman Department of Chemistry 

(1983) Indiana University 

Bloomington, Indiana 47405 

Mark Reshkin Department of Geology 

(1981) Indiana University Northwest 

Gary, Indiana 46408 
Gary E. Dolph Department of Botany 

(1983) Indiana University at Kokomo 

Kokomo, Indiana 46901 

Alice S. Bennett Department of Biology 

(1984) Ball State University 

Muncie, Indiana 47306 

♦President and Secretary serve as ex-officio members of the Research Grants 
Committee. 



STANDING COMMITTEES APPOINTED BY THE PRESIDENT 
(The President and President-Elect are ex-officio members of all committees) 

Academy Representative to AAAS 
Section and Delegate to NAAS 

Earl A. Holmes Department of Biology 

(1980) St. Mary's College 

Notre Dame, Indiana 46556 

Auditing Committee 

William N. Doemel, Chairman Department of Biology 

Wabash College 
Crawfordsville, Indiana 47933 

Eugene P. Schwartz Department of Chemistry 

DePauw University 
Greencastle, Indiana 46135 

Youth Activities Committee 

Donald R. Winslow, Chairman Department of Science and Environ- 
mental Education 
Indiana University 
Bloomington, Indiana 47405 

Keith Hunnings, Assistant 

Chairman and Co-Director, 

Junior Academy of Science Science Department 

New Haven High School 

1300 Green Road 

New Haven, Indiana 46774 

Leota Skirvin, Assistant 

Chairman and Co-Director, 

Junior Academy of Science Brown County High School 

P. 0. Box 68 

Nashville, Indiana 47448 



Indiana Academy of Science 



Karl L. Kaufman, Assistant 
Chairman and Director, 
Youth Science Development 



Department of Mental Health 
5 Indiana Square 
Indianapolis, Indiana 46204 



Walter A. Cory, Jr., Assistant 
Chairman and Director, 
Indiana Science Talent Search 



. . Coordinator for School Science 
Memorial Hall West 108 
Indiana University 
Bloomington, Indiana 47405 

Lloyd Anderson, Assistant 
Chairman and Director of 

Science Fairs Indianapolis Newspapers 

Educational Services Coordinator 
307 North Pennsylvania Avenue 
Indianapolis, Indiana 46206 

F. Keith Ault Department of Chemistry 

Ball State University 
Muncie, Indiana 47306 

Kenneth Bush West Lafayette Senior High School 

Grant and Leslie Avenue 
West Lafayette, Indiana 47906 

Jerry M. Colglazier Office of State Superintendent of 

Public Instruction 
State House 229 
Indianapolis, Indiana 46204 

John C. Moody Division of Education 

Indiana University Southeast 

4201 Grant Line Road 

P. 0. Box 679 

New Albany, Indiana 47150 

Virginia Rhodes East Noble High School 

Kendallville, Indiana 46755 

Jane Tucker St. John the Baptist School 

4500 Fairfield Avenue 
Fort Wayne, Indiana 46807 

Ray Weatherholt Floyd Central High School 

R. R. 2, Box 445 A 

New Albany, Indiana 47150 

Library Committee 

Lois Burton, Chairman Indiana State Library 

Indianapolis, Indiana 46204 

William A. Daily 5884 Compton Street 

Indianapolis, Indiana 46220 

John F. Pelton Department of Botany 

Butler University 
Indianapolis, Indiana 46208 



Officers and Committees 

James A.Clark Assistant State Entomologist 

5519 East 21st Street 
Indianapolis, Indiana 46218 

Nominations Committee 

William B. Hopp, Chairman Department of Life Sciences 

Indiana State University 
Terre Haute, Indiana 47809 

Betty D. Allamong Department of Biology 

Ball State University 
Muncie, Indiana 47306 

Donald J. Cook Department of Chemistry 

DePauw University 
Greencastle, Indiana 46135 

Program Committee 

Duvall A. Jones, Co-Chairman Biology Department 

St. Joseph's College 
Rensselaer, Indiana 47978 

Andrew G.Mehall, Co-Chairman Biology Department 

St. Joseph's College 
Rensselaer, Indiana 47978 

Publications Committee 

William R. Eberly, Chairman Department of Zoology 

Manchester College 

North Manchester, Indiana 46962 

William B. Bunger Department of Chemistry 

Indiana State University 
Terre Haute, Indiana 47809 

Lois Burton Indiana State Library 

Indianapolis, Indiana 46204 

Walter A. Cory, Jr Coordinator for School Science 

Memorial Hall West 108 
Indiana University 
Bloomington, Indiana 47405 

Clyde W. Hibbs Department of Natural Resources 

Ball State University 
Muncie, Indiana 47306 

Marion T. Jackson .Department of Life Sciences 

Indiana State University 
Terre Haute, Indiana 47809 

Paul C. MacMillan Department of Biology 

Hanover College 
Hanover, Indiana 47243 

Wilton N. Melhorn Department of Geosciences 

Purdue University 

West Lafayette, Indiana 47907 

Elmer Nussbaum Department of Physics 

Taylor University 
Upland, Indiana 46989 



10 



Indiana Academy of Science 



Benjamin Moulton 



Gertrude L. Ward 



Department of Geography and 

Geology 
Indiana State University 
Terre Haute, Indiana 47809 
Department of Biology 
Earlham College 
Richmond, Indiana 47374 



Science and Society Committee 

William Beranek, Jr., Chairman Holcomb Research Institute 

(1981) Butler University 

Indianapolis, Indiana 46208 

Clarence Dineen Department of Biology 

(1980) St. Mary's College 

Notre Dame, Indiana 46556 

Charles E. Wier AMAX Coal Company 

(1980) 105 South Meridian Street 

Indianapolis, Indiana 46225 
Howard R. Youse Department of Botany 

(1980) DePauw University 

Greencastle, Indiana 46135 

Stanley L. Burden Department of Chemistry 

(1981) Taylor University 

Upland, Indiana 46989 

Walter Cory, Jr Coordinator for School Science 

(1981) Memorial Hall West 108 

Indiana University 
Bloomington, Indiana 47405 

Robert Menke St. Henry Road 

(1981) Huntingburg, Indiana 47542 

Clyde R. Metz Department of Chemistry 

(1981) Indiana University-Purdue University 

1201 East 38th Street 
Indianapolis, Indiana 46204 

Craig E. Nelson Department of Biology 

(1981) Indiana University 

Bloomington, Indiana 47405 

Jerry J. Nisbet Department of Biology 

(1981) Ball State University 

Muncie, Indiana 47306 
John B. Patton Indiana Geological Survey 

(1981) 611 North Walnut Grove 

Bloomington, Indiana 47405 

Donald J. Cook Department of Chemistry 

(1982) DePauw University 

Greencastle, Indiana 46135 

Willis H. Johnson Department of Biology 

(1982) Wabash College 

Crawfordsville, Indiana 47933 



Officers and Committees 



11 



James M. Shuler Director 

(1982) Office of Chemical Risk 

Indiana State Board of Health 
1330 West Michigan Street 
Indianapolis, Indiana 46206 

Membership Committee 

Ralph A. Llewellyn, Chairman Department of Physics 

Indiana State University 
Terre Haute, Indiana 47809 

Robert H. Cooper, 

Emeritus Membership Department of Biology 

Ball State University 
Muncie, Indiana 47306 

Walter A. Cory, Jr Coordinator for School Science 

Memorial Hall West 108 
Indiana University 
Bloomington, Indiana 47405 

Jackson L. Marr Department of Biology 

Indiana State University-Evansville 
Evansville, Indiana 47712 

Donald R. Winslow, 

Club Membership Department of Science and Environ- 
mental Education 
Indiana University 
Bloomington, Indiana 47405 

Fellows Committee 

Richard L. Conklin (1980) 

Chairman Department of Physics 

Hanover College 
Hanover, Indiana 47243 

Paul H. Gebhard (1980) Institute for Sex Research 

Indiana University 
Bloomington, Indiana 47405 

Russell K. Stivers (1980) Department of Agronomy 

Purdue University 

West Lafayette, Indiana 47907 

Eugene D. Weinberg (1980) Department of Microbiology 

Indiana University 
Bloomington, Indiana 47405 

Wilton N. Melhorn (1981) Department of Geosciences 

Purdue University 

West Lafayette, Indiana 47907 

B. Elwood Montgomery (1981) Department of Entomology 

Purdue University 

West Lafayette, Indiana 47907 

John F. Pelton (1981) Department of Botany 

Butler University 
Indianapolis, Indiana 46208 



12 Indiana Academy of Science 

William W. Bloom (1982) Department of Biology 

Valparaiso University 
Valparaiso, Indiana 46383 

John J. Favinger (1982) 640 Parkway 

Whiteland, Indiana 46184 

James C. List (1982) Department of Biology 

Ball State University 
Muncie, Indiana 47206 

William G. Kessel (1982) Department of Chemistry 

Indiana State University 
Terre Haute, Indiana 47809 

Resolutions Committee 

Howard R. Youse, Chairman Department of Botany 

DePauw University 
Greencastle, Indiana 46135 

Thomas R. Mertens Department of Biology 

Ball State University 
Muncie, Indiana 47306 

Invitations Committee 

Philip A. St. John, Chairman Department of Zoology 

Butler University 
Indianapolis, Indiana 46208 

Gary C. Steinhardt Department of Agronomy 

Purdue University 

West Lafayette, Indiana 47907 

Necrologist 

Fay K. Daily 5884 Compton Street 

Indianapolis, Indiana 46220 

Parliamentarian 

Clarence Dineen Department of Biology 

St. Mary's College 

Notre Dame, Indiana 46556 



SPECIAL COMMITTEES APPOINTED BY THE PRESIDENT 

Biological Survey Committee 

Theodore Crovello, Chairman Department of Biology 

University of Notre Dame 
Notre Dame, Indiana 46556 

John A. Bacone Director 

Division of Nature Preserves 
Indiana Department of Natural 

Resources 
601 State Office Building 
Indianapolis, Indiana 46204 



Officers and Committees 13 

William B. Crankshaw Department of Biology 

Ball State University 
Muncie, Indiana 47306 

James R. Gammon Department of Zoology 

DePauw University 
Greencastle, Indiana 46135 

Jack R. Munsee Department of Life Sciences 

Indiana State University 
Terre Haute, Indiana 47809 

George R. Parker Department of Forestry and 

Conservation 
Purdue University 
West Lafayette, Indiana 47907 

Victor Riemenschneider Department of Biology 

Indiana University at South Bend 
South Bend, Indiana 46615 

John 0. Whitaker, Jr Department of Life Sciences 

Indiana State University 
Terre Haute, Indiana 47809 

Harmon P. Weeks Department of Forestry and 

Conservation 
Purdue University 
West Lafayette, Indiana 47907 

Frank N. Young, Jr Department of Zoology 

Indiana University 
Bloomington, Indiana 47405 

Preservation of Natural Areas Committee 

Marion T. Jackson, Chairman Department of Life Sciences 

(1981) Indiana State University 

Terre Haute, Indiana 47809 

William R. Orr Department of Geography and Geology 

(1980) Ball State University 

Muncie, Indiana 47306 

George R. Parker Department of Forestry and 

(1980) Conservation 

Purdue University 

West Lafayette, Indiana 47907 

CarlH.Krekeler Department of Biology 

(1981) Valparaiso University 

Valparaiso, Indiana 46383 

Victor Riemenschneider Department of Biology 

(1981) Indiana University at South Bend 

South Bend, Indiana 46615 

John A. Bacone Director 

(1982) Division of Nature Preserves 

Indiana Department of Natural 

Resources 
601 State Office Building 
Indianapolis, Indiana 46204 



14 Indiana Academy of Science 

Robert C. Weber 3649 Algonquin Pass 

(1982) Fort Wayne, Indiana 46809 

Damian V. Schmelz Department of Biology 

(1982) St. Meinrad College 

St. Meinrad, Indiana 47577 

Winona H. Welch DePauw University 

(Honorary) Greencastle, Indiana 46135 

Carrolle Markle Ashfield, Massachusetts 01330 

(Honorary) 
Newsletter Editor 

Walter A. Cory, Jr Coordinator for School Science 

Memorial Hall West 108 
Indiana University 
Bloomington, Indiana 47405 

"Speaker of the Year" Selection Committee 

John B.Patton, Chairman Indiana Geological Survey 

(1980) 611 North Walnut Grove 

Bloomington, Indiana 47405 
Norman J. Norton Department of Biology 

(1980) Ball State University 

Muncie, Indiana 47306 

Robert E. Hale Department of Physics 

(1981) Huntington College 

Huntington, Indiana 46750 

Damian V. Schmelz Department of Biology 

(1982) St. Meinrad College 

St. Meinrad, Indiana 47577 

Academy Representative on Indiana Natural Resources Commission 

Damian V. Schmelz Department of Biology 

St. Meinrad College 

St. Meinrad, Indiana 47577 

Emeritus Member Selection Committee 

Robert H.Cooper, Chairman R. R. 9, Box 242 

Muncie, Indiana 47302 

Winona H.Welch DePauw University 

Greencastle, Indiana 47135 

Harry G. Day Department of Chemistry 

Indiana University 
Bloomington, Indiana 47405 

Howard H.Michaud 301 East Stadium Drive 

Lafayette, Indiana 47906 

SPECIAL APPOINTMENTS OF THE YOUTH ACTIVITIES COMMITTEE 

Indiana Science Talent Search Committee 

Walter A. Cory, Jr., Director Coordinator for School Science 

Memorial Hall West 108 
Indiana University 
Bloomington, Indiana 47405 



Officers and Committees 15 

Mark Bambenek Department of Chemistry 

St. Mary's College 

Notre Dame, Indiana 46556 

Robert L. Henry Department of Physics 

Wabash College 
Crawfordsville, Indiana 47933 

Alfred Schmidt Department of Mathematics 

Rose-Hulman Institute 
Terre Haute, Indiana 47803 

Howard Youse Department of Botany 

DePauw University 
Greencastle, Indiana 46135 

Harold Zimmack Department of Zoology 

Ball State University 
Muncie, Indiana 47306 

Editorial Committee for Proceedings 

Benjamin Moulton, Chairman Indiana State University 

and Editor 

Rita Barr (1979-80) Purdue University 

Charlotte Boener (1979-80) Indiana State University 

Ernest Campaigne (1979-80) Indiana University 

James Gammon (1979-80) DePauw University 

John Pelton (1979-80) Butler University 

Carl Sartain (1979-80) Indiana State University 

B. K. Swartz, Jr. (1979-80) Ball State University 



Indiana Academy of Science 
Spring Meeting 

Minutes of the Executive Committee Meeting 
April 25, 1980 

The meeting was called to order by President Robert E. Henderson at 4:12 
p.m. at Geneva Center, Rochester, IN. 

TREASURER'S REPORT 
Treasurer John A. Ricketts presented a brief report on finances for the 
period January 1, 1980 to April 17, 1980, a summary of which follows: 





Academy 
Accounts 


Administered 
Accounts 


Total 


Balance, January 1, 1980 


$ 4,861.86 


$13,479.32 
6,186.00 
3,256.09 


$18,341.18 


1980 Income 

1980 Expenditures .... 


7,551.52 
10,680.60 


13,737.52 
13,936.69 


Balance, April 17, 1980 


1,732.78 


16,409.23 


18,142.01 









The Treasurer's report was approved. 

ELECTED COMMITTEE REPORTS 
Academy Foundation Committee 

William A. Daily, Chairman, reported Foundation accounts as of March 31, 
1980. The value of the John S. Wright Invested Income Account was $54,000 
and the market values of the Foundation Account and John S. Wright Fund were 
$22,423 and $579,928, respectively. The combined assets represent an increase of 
approximately 4% over those of last year. 

Research Grants Committee 

Ernest E. Campaigne, Chairman, reported that 17 proposals were received; 5 
were funded in full and 9 were partially funded. One proposal was still to be con- 
sidered. A total of $6116 was awarded. Deadline for receipt of proposals for the 
Fall 1980 round of grants is September 15, 1980. 

PRESIDENTIAL APPOINTIVE COMMITTEE REPORTS 
Academy Representative to AAAS and A AS 

Earl A. Holmes, Chairman, reported on the San Francisco meeting of 
January 3-8, 1980 and current activities of Section X, including new rules for ap- 
plication of grant monies. Also announced: The Junior Academy can nominate 
four individuals for subscriptions to Science 80. 

In other business, President Henderson stated that he had a communication 
from AAAS requesting a membership list for advertising Science 80. After some 
discussion, the following motion was presented: 

Motion: That the request from AAAS for Academy membership lists be 
denied. Seconded and carried. 

16 



Minutes of the Executive Committee 17 

Youth Activities Committee 

President Henderson discussed the Science Education Fund and named Karl 
L. Kaufman as Assistant Chairman and Director, Youth Science Development, 
and Lloyd Anderson as Assistant Chairman and Director of Science Fairs. 

In place of Donald R. Winslow, Chairman of Youth Activities Committee, 
President Henderson discussed the concept that the Academy award membership 
in the Academy to approximately 12 high schools whose students were winners in 
the Talent Search. 

Motion: That the Academy award a one-year student membership to the 
winners of the Talent Search and provide a certificate of member- 
ship suitable for framing. Seconded and passed unanimously. 

Biological Survey Committee 

Theodore Crovello, Chairman, discussed the progress on the Computer Data 
Base. The computer files are updated for 1969-78. 

The production of vegetation maps for the State of Indiana was discussed: ap- 
proximately $30,000 is required as a budget to print 1500 maps. Three maps could 
be produced, including the current use map. Ways of obtaining the required 
money were discussed. 

Editor's Report 

Benjamin Moulton, Editor, briefly discussed the printing of the most recent 
Proceedings, Volume 89. 

Program Committee 

Duvall A. Jones, Co-Chairman, commented on preparations for the meeting 
at Saint Joseph's College. Discussion centered on the possibility of having two 
days of meetings for the Fall Program. A main item of concern was how to main- 
tain good attendance on both days. The following trial program schedule was 
agreed by consensus to be tried for this year: 

Two-day meeting, with Executive Meeting Thursday Evening. 

Friday: 

Division Sessions for both Junior Academy and Academy presentations on 
Friday, with expansion into Saturday morning if necessary for overflow 
papers. 

General Business Meeting after Luncheon on Friday. 
Social Hour between 5 and 6 Friday. 

Dinner at approximately 6:00, followed by Annual Banquet and Committee 
meetings. 
Saturday: 

Division Sessions early morning 8-9:45. 

Two parallel symposia to be arranged 10-noon. 

Luncheon 

Lecture-of-the-Year by Dr. Harry Day. 

The meeting was adjourned at 5:52 p.m. 

Respectfully submitted, 

John H. Meiser 

Secretary 



SPRING MEETING 

Minutes of the General Session 

April 25, 1980 

The meeting was called to order by President Robert E. Henderson at 7:32 
p.m. at Geneva Center, Rochester, Indiana. 

President Henderson introduced Father Damian V. Schmelz as Speaker of 
the Year. Father Schmelz presented a stimulating slide-illustrated lecture en- 
titled "Stewardship of Natural Resources in the State of Indiana." 

At 8:55 p.m. President Henderson reconvened the general business meeting. 
Announcements were made concerning the morning field trips and workshop. 

Dr. Ernest Campaigne, Chairman, Research Grants Committee, announced 
that Academy grant forms were available through him. 

Section chairmen were to meet at end of the meeting to discuss the Fall 
meeting. 

Meeting adjourned at 9:07 p.m. 

This meeting was followed by an interesting discussion and slide show on 
"Mushrooms in Indiana," presented by John C. Tacoma. 

Respectfully submitted, 
John H. Meiser 
Secretary 



Indiana Academy of Science 
96th ANNUAL FALL MEETING 

Minutes of the Executive Committee Meeting 

November 6, 1980 

The meeting was called to order by Treasurer John A. Ricketts at 7:58 p.m. 
in Raleigh Hall Conference Room at Saint Joseph's College, Rensselaer, Indiana. 
The minutes of the Executive Committee and of the General Session of the Spring 
1980 meeting at Geneva Center were read and approved. 

TREASURER'S REPORT 
Treasurer John A. Ricketts presented the Academy financial report for the 
period January 1, 1980 through October 31, 1980. A brief summary follows: 

Academy Administered 

Accounts Accounts Totals 

Balance, January 1,1980 $ 4,861.86 $13,479.32 $18,341.18 

Income 14,324.77 35,521.38 49,846.15 

Expenditures 17,763.26 31,354.04 49,121.30 

Balance, October 31, 1980 $ 1,423.37 $17,646.66 19,070.03 

There were 841 paid members as of October 31, 1980, including 23 sustaining 
members, 488 senior members, 51 student members, and 52 new members. The 
Treasurer's report was approved. 

ELECTED COMMITTEE REPORTS 
Academy Foundation Committee 

William A. Daily, Chairman, reported Foundation accounts as of September 30, 
1980. A summary of that report follows: 
I. Foundation Account 

Balance on September 30, 1979 $ 232.00 

Income as of September 30, 1980 1,829.97 

Disbursements to Academy -300.00 

Cash balance as of September 30, 1980 1,761.97 

Total Market Value $23,948.00 



II. JohnS. Wright Fund 

Total income 10/1/79 to 9/30/80 
No disbursements 

Total Market Value 



$27,891.93 



$595,929.80 



III. JohnS. Wright Investment Income Account 
Total interest 10/1/79 to 9/30/80 
Disbursements from account: 

Research Grants 

Publication of IAS Proceedings 

Indiana National Bank Fee 



$ 6,447.48 

11,187.00 

11,906.63 

2,636.00 



The Academy portfolio of the John S. Wright Fund account, Mr. Daily noted, 

19 



20 Indiana Academy of Science 

has decreased in market value $27,697 or 4.4%, due to adverse market conditions. 
However, the Investment Income Account has increased to $57,000, yielding about 
12%. 

Research Grants Committee 

Ernest Campaigne, Chairman, reported that 14 papers, representing contribu- 
tions from 7 institutions, had been received for the Fall '80 competition. Of these, 2 
received full funding and 9 partial funding. For the year, 31 proposals were received, 
with 7 receiving full funding and 19 partial funding, with an average award of $444. 
The total expenditure was $11,090. 

In addition, eight small grants jointly sponsored by the Academy and AAAS 
were awarded to Secondary School Student applicants for a total of $690. 

Discussion concentrated on the accountability of the researchers to report their 
progress to the Grants Committee. 

Bonding Committee. No report. 
Academy Representative to NAAS 

President Robert E. Henderson reported that he would represent the Academy 
at the AAAS meeting in Toronto, at which meeting he would present a paper. 

Youth Activities Committee 

Donald R. Winslow, Chairman, discussed the program for the ISTA workshop, 
"How to Make Classroom Science Exciting." He reported that the workshop was well 
received. 

Walter A. Cory, Jr., Assistant Chairman and Director of the Indiana Science 
Talent Search, reported that brochures have been distributed to each high school 
encouraging participation in the Science Talent Search. This year again Kappa 
Kappa Kappa will support this work with a $2,000 grant. 

Keith Hunnings, Assistant Chairman and Co-Director of the Junior Academy 
of Science, reported that there are now 30 science clubs that are active. The conti- 
nuing growth was related to the dedication of the teachers and the Friday 
meetings of the Junior Academy. 

Discussion ensued concerning whether the best paper or papers from the 
Junior Academy could be presented before the Academy. 

Motion; That the Outstanding Junior Academy paper be presented at the 
proper forum in the Senior Academy. 
Seconded and passed. 

Lloyd Anderson, Assistant Chairman and Director of Science Fairs, reported 
that under the auspices of the IAS 30,000 students participate in the Science and 
Engineering Fairs. Indiana is unique in its organization and support of such fairs. 
He also reported that 60 people, including the two student winners in each of the 
12 Regional Science and Engineering Fairs, will travel to Milwaukee for the Inter- 
national Science and Engineering Fair. The Indiana Science Education Fund will 
subsidize approximately $10,000 of the $19,900 cost. 

He also noted that the rules governing the types of research are becoming 
more stringent. 

Library Committee 

William A. Daily, reporting for Lois Burton, Chairperson, indicated that the 
Proceedings, Volumes 88 and 89, had been distributed. Copies were also sent to 
approximately 500 libraries, institutions, and societies on the exchange list. In ad- 
dition, 43 new titles were received and 120 volumes of the library's journals were 



Minutes of the Executive Committee 21 

prepared for binding. Some 269 interlibrary loan requests were also filled during 
the year. 

Publications Committee 

William R. Eberly, Chairman, reported one monograph is ready for editing. 
The Centennial History of the Academy is now in preparation by William and Fay 
Daily. The committee continues to seek members to prepare additional titles, 
especially an update of the History of Indiana Scientists. 

Science and Society Committee 

Walter A. Cory, Jr., distributed a questionnaire to establish whether a need 
exists to reinstitute the Speakers Bureau. 

Biological Survey Committee 

John A. Bacone, Indiana Department of Natural Resources, reported that 
work was continuing on the Federal Grant for Fish and Wildlife Study and that a 
paper was being presented. 

Membership Committee 

Walter A. Cory, Jr., reported that a new supply of brochures was available 
and that prospective candidates would be sent letters of invitation. 

Fellows for 20 years are being issued special pink name tags as a sign of 
honor at this meeting. 

Fellows Committee 

Richard L. Conklin, Chairman, gave the committee report and moved that 
those named be presented to the membership for election to the rank of Fellow. 
Seconded and passed. 

Emeritus Membership 

Robert H. Cooper presented the names of Dr. L. S. McClung, Department of 
Biology, Indiana University (1940) and Dr. W. Max Stark, Carmel, Indiana (1948) 
as fulfilling the requirements of Emeritus Membership. (Initial membership year 
shown in parentheses.) 

Motion: That the persons presented be elected to Emeritus Membership. 
Seconded and passed. 

"Speaker of the Year" Selection Committee 

John B. Patton, Chairman, announced that Harry G. Day's speech is retitled 
"Progress in Resolving Food Safety Problems: Systematic Evaluation of GRAS 
Food Substances." Patton also indicated participating colleges and universities 
would be offered the opportunity to have Dr. Day, our "Speaker of Year," speak 
at their institutions. 

Program Committee 

Duvall A. Jones, Co-Chairman, indicated some of the detail in the prepara- 
tions for the Fall meeting. 

Nominations Committee 

William B. Hopp, Chairman, submitted the nominations to be presented to 
the general membership. 



22 Indiana Academy of Science 

Motion: To accept the recommendations of the Nominations Committee. 
Seconded and passed. 

NEW BUSINESS 

Donald D. Gray, Chairman of the Engineering Section, inquired to the extent 
that papers submitted to the Proceedings were reviewed. Benjamin Moulton, 
Editor of the Proceedings, spoke of the problems of refereeing the papers. 

Meeting adjourned at 10:05 p.m. 

Respectfully submitted, 
John H. Mesier 
Secretary 



FALL MEETING 

Minutes of the General Session 
November 7, 1980 



The Business Session of the 96th Annual Meeting of the Academy was called 
to order by President Robert E. Henderson at 1:10 p.m. in Science Hall 
Auditorium at Saint Joseph's College, Rensselaer, Indiana. 

Father Charles Banet, President of Saint Joseph's College, welcomed the 
Academy members on behalf of the college. 

President Henderson introduced the Secretary of the Academy who sum- 
marized the committee reports and informed the membership of the official 
actions taken by the Executive Committee the previous evening. 

The Secretary announced that because all divisional election results were not 
available, the list of chairmen and chairmen-elect of the divisions for 1981 would 
not be read but would appear in the next Newsletter and be published in the 
minutes of the meeting. That list follows: 



ANTHROPOLOGY 

Chairman: 



Chairman-Elect: 



BOTANY 



Chairman: 
Chairman-Elect: 

CELL BIOLOGY 

Chairman: 
Chairman-Elect: 

CHEMISTRY 

Chairman: 
Chairman-Elect: 

ECOLOGY 

Chairman: 
Chairman-Elect: 

ENGINEERING 

Chairman: 
Chairman-Elect: 

ENTOMOLOGY 

Chairman: 
Chairman-Elect: 

ENVIRONMENTAL QUALITY 
Chairman: 
Chairman-Elect: 

GEOLOGY AND GEOGRAPHY 
Chairman: 
Chairman-Elect: 



Charles P. Warren 
Curtis H. Tomak 

Charles T. Hammond 
Kathryn Wilson 

Kara W. Eberly 
Douglas W. Duff 

Donald G. Clemens 
Margaret A. Cavanaugh 

Richard W. Greene 
David F. Spencer 

Mark H. Houck 
Warren W. Bowden 

David K. Reed 
Robert Pinger 

David Peterson 
Patrick J. Sullivan 

Henry H. Gray 

Akhtar Husain Siddiqi 



23 



24 Indiana Academy of Science 

HISTORY OF SCIENCE 

Chairman: Everett F. Morris 

Chairman-Elect: Gene Kritsky 

MICROBIOLOGY AND MOLECULAR BIOLOGY 

Chairman: Dorothy Adalis 

Chairman-Elect: Kenneth Brunson 

PHYSICS AND ASTRONOMY 

Chairman: L. Dwight Farringer 

Chairman-Elect: Roger Roth 

PLANT TAXONOMY 

Chairman: Richard J. Jensen 

Chairman-Elect: Don Hendricks 

SCIENCE EDUCATION 

Chairman: Charles L. Gehring 

Chairman-Elect: Patricia A. Zeck 

SOIL AND ATMOSPHERIC SCIENCES 

Chairman: Robert F. Dale 

Chairman-Elect: John T. Snow 

ZOOLOGY 

Chairman: Larry Ganion 

Chairman-Elect: David Sever 

Fay K. Daily presented the Necrologist's report, which included the names of 
three members: 

Max William Gardner 
Karl Stone Means 
Helen E. Reed 

William B. Hopp, Chairman of the Nominations Committee, placed the follow- 
ing slate in nomination: 

President Robert E. Henderson 1981 

President-Elect William R. Eberly 1981 

Editor Donald R. Winslow 

Academy Foundations William A. Daily 

Bonding Committee William B. Bunger 

Research Grants Committee Uwe J. Hensen 

No nominations were made from the floor. The motion was made to accept those 
named by the Nominations Committee as elected. Motion was seconded and passed. 

Richard L. Conklin, Chairman of the Committee on Fellows, moved that 
William B. Bunger 
Donald P. Franzmeier 
Lawrence A. Schaal 
Joseph R. Siefker 
be elected to the rank of Fellow of the Indiana Academy of Science. Seconded and 
carried. Conklin then presented certificates to the newly-elected Fellows. 

Howard Youse, Chairman of the Resolutions Committee, presented the 
following: 



Minutes of the General Session 25 

RESOLUTION: 

WHEREAS: The Indiana Academy of Science is deeply grateful to 
Saint Joseph's College for their invitation to hold its 
96th annual meeting on their campus; and 

WHEREAS: Administration, faculty, and students alike have 
cooperated in providing us their facilities for this 
meeting; be it 

RESOLVED: That the Academy members here assembled express 
their sincere appreciation to Father Charles Banet, 
President of Saint Joseph's College, for all the 
courtesies that have been extended to the Academy 
during this meeting. We are especially grateful to Dr. 
Duvall A. Jones and Dr. Andrew G. Mehall and their 
committee for the arrangements of the entire program 
and the comfort and conveniences provided the 
membership. We also express our sincere thanks to all 
members who organized and participated in the special 
symposia on "Coal and Indiana" and "The Evolu- 
tion/Creation Controversy." 

NEW BUSINESS 

Duvall A. Jones, Co-Chairman, Program Committee, announced the showing 
times of a film provided by Eli Lilly Company entitled "Recombinant Techniques." 

Donald R. Winslow, Editor for 1981, requested that abstracts for the papers 
presented at the meeting be submitted as soon as possible. 

Some discussion followed concerning the location of future meetings. The 
Fall 1981 meeting is being arranged at Wabash College, with the Spring meeting 
at Turkey Run State Park on April 17-18. 

The meeting adjourned at 1:50 p.m. and the presentation of papers continued. 

The annual banquet was held in the Halleck Student Center at 6:30 p.m. with 
President Henderson presiding. After the banquet President Henderson address- 
ed the attendees on "Scientific Research and Economic Indicators." 

The General Session reconvened at 8:45 a.m., November 8 with presented 
papers followed by symposia, 

"The Dangerous Position of the Science Teacher: The Evolution/Creation 

Controversy" and "Coal and Indiana," 
arranged by the Committee on Science and Society, William Beranek, Jr., Chair- 
man. 

Following luncheon in the Halleck Student Center, the Lecture-of-the-Year 
was presented by Harry Day, Professor Emeritus of Chemistry, Indiana Univer- 
sity. An interesting and informative time was had by all who attended. 

Respectfully submitted, 
John H. Mesier 
Secretary 



FINANCIAL REPORT 

January 1-December 31, 1980 





I. ACADEMY ACCOUNTS 










Income 


Budgeted 


Expenditures 


Budgeted 


Dues 


$ 7,724.00 


$ 7,350.00 


$ - 


$ - 


Reprints 


6,532.11 


3,500.00 


7,636.29 


3,250.00 


Interest 


1,214.47 


1,000.00 


- 


- 


Miscellaneous 


1,810.18 


0.00 


15.00 


100.00 


Secretary 


- 


- 


536.60 


500.00 


Treasurer 


- 


- 


308.35 


400.00 


General Office.... 


- 


- 


247.20 


750.00 


Officer Travel 


- 


- 


150.00 


225.00 


Membership Committee 


_ 


- 


100.00 

3,350.00 

0.00 


250.00 


Transfer Administered Accounts 


_ 


3,350.00 


President's Fund 


- 


150.00 


Newsletter 


- 


- 


500.00 


500.00 


Speaker of the Year 


- 


- 


700.00 


700.00 


Program Committee 


_ 


- 


2,560.08 
700.00 


1,500.00 


Editor's Expense 


- 


700.00 


Youth Activities 


_ 


- 


30.30 
1,050.00 


50.00 


Biological Survey Committee 


- 


2,250.00 


A A AS Representative 


- 


- 


395.00 


300.00 


Section Chairman Expenses 




— 


36.00 
700.00 


50.00 


CPA Fees 


_ 


750.00 


Lawyer's Fees 


- 


- 


0.00 
$19,014.82 


100.00 




$17,280.76 


$11,850.00 


$15,875.00 



II. ADMINISTERED ACCOUNTS 





January 1 


1980 


1980 E 


ecember 31 




Balance 


Income 


Expenditures 


Balance 


Junior Academy 


$ 1,454.01 


$ 300.00 


$ 0.00 


$ 1,754.01 


Science Talent Search.. 


303.59 


2,840.00 


2,345.80 


797.79 


Science And Society 


2,734.91 


500.00 

11,990.00 

0.00 


173.42 

11,880.00 

0.00 


3,061.49 


Research Grants 


- 6,447.68 


- 6,337.68 


Lilly Library Fund III 


2,632.76 


2,632.76 


Lilly Library Fund V_._._ 


4,513.20 


0.00 


13.00 


4,500.20 


J.S. Wright Library Fund 


134.28 


0.00 


0.00 


134.28 


Natural Areas 


0.00 


0.00 


0.00 


0.00 


Library Binding 


2,247.30 


1,500.00 


2,264.70 


1,482.60 


Proceedings: Publications 


2,400.89 


12,677.63 


11,845.63 


3,232.89 


Proceedings: Mailing 


707.04 


100.00 


284.29 


522.75 


Publications and Sale of 










Monographs 


2,299.02 


571.00 


0.00 


2,870.02 


Publications: Clerical 


500.00 


0.00 
10,226.75 


500.00 
9,822.11 


0.00 


Dept. of Interior Grant 


0.00 


404.64 


14-16-0003-79-133 












$13,479.32 


$40,705.38 


$39,128.95 


$15,055.75 



III. SUMMARY 



Academy Administered 
Accounts Accounts 



Balance 1/1/80...... 

Income 

Expenditures 

Balance 12/31/80. 



$ 4,861.86 


$13,479.32 


$18,341.18 


17,280.76 


40,705.38 


57,986.14 


19,014.82 


39,128.95 


58,143.77 


3,127.80 


15,055.75 


18,183.55 



26 



Financial Report 



27 



IV. BANK BALANCE 12/31/80 

First Citizens Bank and Trust Co. (Checking Account) 

Savings Accounts: 

C.D.6Mos., First Citizens - 

First Western Savings (matures 6/16/81) 



$ 3,006.40 



10.235.79 

4,941.36 

$18,183.55 



V. SUMMARY OF TRUST FUNDS 

A. Foundation Account (00430-00-0) 

Income Balance (1/1/80) 

Dividends and Interest.... 

Disbursements for 1980: 

Research Grants * 300.00 

Transfer -« 

$ 1,300.00 

Principal Cash 12/31/80 

Investments Carrying Value 

B. John S. Wright Fund (00430-01-9) 

Income Balance (1/1/80) 

Dividends and Interest 

Disbursements for 1980: 

$ 2,636.00 
Fees 

Transfer to (00430-02-08) - 22,513.86 

Account transfer - 203,18 

$25,353.04 

Principal Cash Balance (12/31/80) 

Investments Carrying Value...... 

C. Indiana Academy of Science Invested Income Account (00430-02-08) 

Income Cash Balance (1/1/80) - 

Interest 

Disbursements: 

Account Transfer - $6,463.00 

Principal Cash (1/1/80) 

Investments Value (1/1/80) - 

Receipts for 1980: 

t , 1 cm $49,000.00 

Investments bold.. 

„. ., $22,513.86 

Miscellaneous _ -Z — ! 

$71,513.86 

Disbursements: 

Investments Purchased....... _ - $53.988.b0 

Academy Expenses: 

Research... __ "•«*>■<»> 

Printing Proceedings 11.096.63 

$76,985.23 

, „ ( $ 6.463.00 

Account Transfer - 

Principal Cash Balance (12/31/80) 

Investments Carrying Value 



$ 567.81 
1,855.52 



- 1,300.00 
$ 1,123.33 



$ 1,530.95 
28,398.92 



- 25.353.04 



53.40 
27.178.47 



$ 4,576.83 






$ 754.03 




$320,017.47 


$ 810.33 




7,153.90 




- 6.463.00 





$ 1,501.23 

0.00 

49.000.00 



$ 991.63 
53.988.60 



VI. NOTES 

Membership for 1980: 

1.002 active members (25 sustaining. 522 senior. 287 regular. 60 student. 101 ementus) 
69 new members 
78 members dropped for non-payment of 1980 dues. 



28 Indiana Academy of Science 

Dues Structure for 1980: 

$ 2.00 for student memberships 

5.00 for regular memberships and club memberships 
10.00 for senior memberships 
25.00 for sustaining memberships 
2.00 additional for family memberships 
1.00 additional for initiation or reinstatement fee. 

Special memberships include: Life membership ($300.00), Corporate memberships (150-$500.00), Institu- 
tional memberships ($50-$150.00) 

Reprints: 

Reprint charges for Volumes 88 and 89 were $7,636.29; reprint charges collected from authors amounted 
to $6,532.11. 

Research Grants: 

$11,880.00 was awarded to the following persons to support their research activities: William Bandy 
(Indiana U.), John Fezy (U. of Notre Dame), James Vogelman (Indiana U.), Thomas Jones (Purdue U.), 
David Osgood (Butler U.), Kathleen Horwath (U. of Notre Dame), Donald French (Indiana U.), Carol 
Summers (Indiana U.), Mary Richeson (Purdue U.), Larry Hauser (U. of Notre Dame), Robert Rogers 
(Purdue U.), Joseph Tatarewicz (Indiana U.), Ernest Shull (Manchester College), Mark Loyd (Ball State 
U.), Betty Allamong (Ball State U.), Thad Godish (Ball State U.), Donald Hudson (Indiana U.), Steven 
Manchester (Indiana U.), Joanne Payton (U. of Notre Dame), Walter Pierce (Ball State U.), David 
Spencer (IUPUI), Patrick Sullivan (U. of Notre Dame), Clifford Teme (Purdue U.), Jerry Nisbet (Ball 
State U.), Frederick Woodson (Indiana State U.), Rick Meyer (East Noble High School), Dana Kincaid 
(Heritage Christian School), Eleanor Pollak (John Adams High School), Tom Sedgewick (East Noble 
High School), Tom Buether (Marquette High School) Mark Paulik (Marquette High School), Richard 
Bush (Marquette High School), and Paul Worthington (Marquette High School). 

Publications: 

Sales during 1980 amounted to $571.00. The cost of publishing Volume 88 and 89 amounted to 
$11,845.63 (Academy's share). 

VII. BUDGET FOR 1981 

The following budget was approved by the Budget Committee on November 29, 1980. 

Academy Accounts 

Anticipated Income: 

Dues $ 7,500.00 

Reprint Charges to Authors 4,000.00 

Interest _ _ 1,200.00 

Miscellaneous __ ___ ___ 0.00 

$12,700.00 

Budgeted Expenditures: 

Publishing Charges for Reprints $ 3,500.00 

Secretary.. _ 500.00 

Treasurer _ ___ 400.00 

General Office _____ _. 350.00 

Officer's Travel _ _ 225.00 

Membership 250.00 

President's Fund 150.00 

Newsletter __ 650.00 

Speaker of the Year 700.00 

Program Committee _ 1,700.00 

Editor's Expenses 900.00 

Youth Activities _ _ ____ 250.00 

Biological Survey Committee _.._ __._ 2,250.00 

AAAS 0.00 

Public Relations 100.00 

Section Chairman Expenses _____ _ 50.00 

CPAFee_ 750.00 

Lawyer's Fee 100.00 

Miscellaneous 100.00 

Transfer from Administered Accounts ( - 1,200.00 ) 

$11,725.00 



Financial Report 29 

Administered Accounts: 

Junior Academy $ 0.00 

Science Talent Search 0.00 

Science and Society ( - 1,000.00) 

Library Binding 1,500.00 

Proceedings: Postage 300.00 

Proceedings: Publication ( - 2,000.00) 

$(-1,200.00) 

Endowment Funds 

Anticipated Income: 

IAS Foundation $ 1,000.00 

J.S. Wright Invested Income „ _ 30,000.00 

Invested Income Carry-Over _. 57,000.00 

$88,000.00 

Expenditures: 

Bank Fee _ $ 2,750.00 

Reserach Grants (including $300.00 from IAS Foundation) 15,000.00 

Publication of Proceedings (Academy's share) 6,250.00 

Publications: Special _ 5,000.00 

$29,000.00 

Restricted Accounts 
Anticipated Income 

Science Talent Search from Tri Kappa $ 2,500.00 

Department Interior Grant 14-16-0003-79-133 3,500.00 

$ 6,000.00 

Expenditures: 

Science Talent Search $ 2,500.00 

Department Interior Grant 14-16-0003-79-133.... 3,500.00 

$ 6,000.00 

Respectfully submitted, 
John A. Ricketts, Treasurer 



INDIANA JUNIOR ACADEMY OF SCIENCE 

OFFICERS 

President Jamie Sibbitt, Paoli, Jr.-Sr. High School 

President Elect Ernie Tseng, Highland High School 

Secretary Jody Sibbitt, Paoli, Jr.-Sr. High School 

JUNIOR ACADEMY COUNCIL 

Larry Bennet (Co-Chair, Polemic) Burris Jr. High School, Mitchell 

Michael Kobe (Chair, Paper Planes) Gavit High School, Hammond 

Cheryl Mason (Co-Chair, Polemic) Highland High School, Highland 

Thomas Anderson (Chair, Junior Paoli Jr.-Sr. High School, 

Scientist) Paoli 

Carrol Ritter (Chair, Junior Paoli Jr.-Sr. High School, 

Scientist) Paoli 

Jane Tucker (Chair, Senior Papers) Saint John the Baptist, Ft. Wayne 

Leota Skirvin (Chair, Junior Brown County High School, 

Papers; IJAS Co-Director) Nashville 

Keith Hunnings (Chair, Science New Haven High School, 

Fiction; IJAS Co-Director) New Haven 

Donald Winslow (Youth Council- Indiana University, Bloomington 

Director) 

PROGRAM 

FORTY-EIGHTH ANNUAL MEETING 

November 7, 1980 

Saint Joseph's College, Rensselaer, Indiana 

8:30-11:00 AM Registration, Halleck Student Center 

9:00- 9:30 AM Welcome by Father Charles Banet, President of Saint Joseph's 
College. 

Introduction of candidates and announcements. Ballroom, 
Halleck Student Center. 
9:35-11:00 AM Papers Presented, Halleck Student Center, Second Floor. 
9:35-10:30 AM Paper Planes. Ballroom, Halleck Student Center. 
9:35- AM Junior Scientist Interviews. Halleck Student Center. North 

Lounge. 
10:15-10:55 AM Group "A" Polemic. Ballroom, Halleck Student Center. 
11:00-11:40 AM Group "B" Polemic. Ballroom, Halleck Student Center. 
12:00- 1:00 AM Lunch. Cafeteria, Halleck Student Center. 



Afternoon 
1:30 PM 

2:45 PM 



Awards presented. Ballroom, Halleck Student Center. 
Adjournment. 



CANDIDATES FOR STATE OFFICE 1981-82 
President Elect: Bourdon, Michael Bryan-Marquette High School 

Sibbitt, Jody Lynn-Paoli Jr.-Sr. High School 
Secretary: Karr, Rick G.-Highland Sr. High School 

Pardus, Philip Anthony-Pierce Jr. High School 



30 



Junior Academy Report 31 

SENIOR DIVISION PAPERS 
HALLECK STUDENT CENTER (2nd Floor) 
LIFE SCIENCE 
9:35- 9:45 Kelli Copeland-East Noble High School 

"The Effects of Oligodynamics on Staphylococcus aureus and 
Escherichia coli" 
9:50-10:00 Ernie Tseng-Highland High School 

"Acid Phosphatase." 
10:05-10:15 Tom Sedgwick-East Noble High School 

"Crown Gall: A Study of Botanical Cancer and Its Control." 
10:20-10:30 Jamie Sibbitt-Paoli Jr.-Sr. High School 

"Cigarette Smoke Residue Quantity Comparisons: The 
Effects on Bacterial Survival." 
10:35-10:45 Luralynn Ustanik-Highland High School 

"Studies on the Co-aggregation of Steptococcus sanguis and 
Actinomyces naeslundii." 
10:50-11:00 Mark A. Paulik-Marquette High School 

"Laetrile Cancer Research: A Complete Metabolic Program 
Consisting of an Anti-Tumor Regimen of Laetrile, Vitamin A, 
and Retenzyme, on Mammalian Lung Cancer." 
11:05-11:15 Michelle Trinkle-Paoli Jr.-Sr. High School 

"The Absorption, Distribution, and Utilization of Water and 
Minerals by Various Herbaceous Plants Using Radioisotopes 
as Tracers." 



SENIOR DIVISION PAPERS 
HALLECK STUDENT CENTER (2nd Floor) 
PHYSICAL SCIENCE 
9:35- 9:45 Michael A. Warot-Gavit Jr.-Sr. High School 

"The Use of Pulse Width Modulation for Stereo Data Trans- 
mission." 
9:50-10:00 Thomas Beutner-Marquette High School 

"Wind Tunnel Design and Remote Piloted Testing of Multi- 
Bladed Rotor System, Experimental Airfoils, and Wing- 
lets as High Performance Devices on Rotary and Stationary 
Wing Aircraft." 
11:50-12:05 Douglas Ayers-Brown County High School 

"Laser and Reflection." 
10:20-10:30 Dan Gehring-New Haven High School 

"Translation of German to English via Computer." 



INTERVIEWS FOR OUTSTANDING JUNIOR SCIENTIST 

SENIOR DIVISION 

North Lounge H.S.C. (1st Floor) 

9:35- 9:50 Ernie Tseng-Highland High School 

9:55-10:10 Jamie Sibbitt-Paoli Jr.-Sr. High School 

10:15-10:30 Thomas Beutner-Marquette High School 



32 Indiana Academy of Science 

JUNIOR DIVISION PAPERS 
HALLECK STUDENT CENTER (2nd Floor) 

9:35- 9:45 Ali Raheem-Marquette (Junior High) School 

"Lung Cancer." 
9:50-10:00 Rick G. Karr-Highland Senior High School 

"Acid Rain, Its Effects on a Small Pond Ecosystem." 
10:05-10:15 Chris Lauer-St. John The Baptist 

"Logic Fun in Math." 
10:20-10:30 Brad Shrader-Pierce Junior High School 

"Pacemakers." 
10:35-10:45 Lauri Spindler-The Canterbury School 

"Chemical Differences in the Water of the Fort Wayne 

Metropolitan Areas as a Function of pH Conductivity, 

Chloride, Hardness and Alkalinity." 
10:50-11:00 James Barsic-Marquette Jr. High School 

"The Design, Comparison, and Construction of Two Basic 

Types of Cloud Chambers-Phase I." 
11:05-11:15 Amy Knight-St. John The Baptist 

"The Formation of Taste." 
11:20-11:30 John Terrill-The Canterbury School 

"An Inductive Analysis of the Possibility of Acid Rain in 

the Fort Wayne Metropolitan Area." 
11:35-11:45 Philip Pardus-Pierce Jr. High School 

"Interferon: Wonder Drug of Modern Medicine." 

INTERVIEWS FOR OUTSTANDING JUNIOR SCIENTIST 

JUNIOR DIVISION 

North Lounge H.S.C (1st Floor) 

10:35-10:45 Rick Karr-Highland Senior High School 

10:50-11:00 Ali Raheem-Marquette Junior High School 

SCIENCE FICTION STORIES 
SENIOR DIVISION 

Richard A. Dahm-Bishop Luers High School 

"You Can't Keep a Good Man Down" 
Tammy Minor-Brown County High School 

"Double Jeopardy" 
Adam Adams-Gavit Jr.-Sr. High School 

"Challenge of the Headless Baseball Team" 
Barry Driver-Gavit Jr.-Sr. High School 

"The Battle of Armageddon" 
Kim Earman-Highland High School 

"Earth to Earth . . . Dust to Dust" 
Kelly Hanson-Highland High School 

"The Final Generation" 
Beth England-Marquette High School 

"Existing In Time" 
J. Genschoreck-Marquette High School 

". . . An In This Corner. . ." 



Junior Academy Report 33 

JUNIOR DIVISION 

Lisa Hill-St. John the Baptist Catholic School 

"Just One Boy" 
Ryan Hill-St. John the Baptist Catholic School 

"Gamma World" 
Joff Giltz-Pierce Junior High School 

"The Flammable Object" 
Paul Tomak-Pierce Junior High School 

"The Desert Devil" 
James Barsic-Marquette High School 

"The Scattering" 
Amer Marsh-Marquette High School 

"A Mouse's Dream Come True" 
Joseph Pawlowski-Highland High School 

"Fatal Mission" 

ACKNOWLEDGEMENT 

The officers, members of the Junior Academy Council and all others con- 
cerned with the program, wish to thank Dr. Duvall A. Jones and Dr. Andrew G. 
Mehall of the Saint Joseph's College Biology Department for arranging the fine 
tours and meeting facilities for our convention. We also want to make special men- 
tion of the time and effort they gave so generously, thereby making our meeting 
such a success. 



Biological Survey Committee Report 

NEW LITERATURE AND WORKS IN PROGRESS 

ABOUT THE BIOTA OF INDIANA 

1980 Annual Report Of The Biological Survey Committee (BSC) 

Of The Indiana Academy Of Science 

BSC Members For 1980: 
Theodore J. Crovello, Chairman, Department of Biology, University of 

Notre Dame, Notre Dame, Indiana 46556, Phone: (219) 283-7496. SUVON: 

Your access code plus 736 + 7496 
John A. Bacone, Indiana Department of Natural Resources 
William Crankshaw, Ball State University 
James R. Gammon, DePauw University 
Jack R. Munsee, Indiana State University, Terre Haute 
George R. Parker, Purdue University, Lafayette 
Victor Riemenschneider, Indiana University at South Bend 
John Whitaker, Jr., Indiana State University, Terre Haute 
Harmon P. Weeks, Purdue University, Lafayette 
Frank N. Young, Jr., Indiana University, Bloomington 

The goal of the Literature Subcommittee of the Academy's Biological Survey 
Committee is to accumulate and maintain published and unpublished references 
on the biota of the State. While in the past the BSC Literature Project has 
presented its findings in conventional printed form, in addition we now 
simultaneously add these contributions to the Indiana Academy's computerized 
literature and works in progress register. It provides update, integrated 
bibliographies on particular topics in response to specific requests. Contact Pro- 
fessor Crovello for further details or to submit literature or project information to 
the data bank. The data bank contains more information about each reference 
than is presented in this printed summary. Items omitted here include: Higher 
taxa (common names) to which organisms belong; Classes or orders of organisms; 
Families; Genera or species; Subjects describing each work; and name, address 
and telephone number of person to contact for further information on each entry. 

The following literature references and works in progress were obtained by 
circulating requests and questionnaires to chairpersons of 69 Life Science depart- 
ments throughout the State, and to participants at the Academy's Fall Meeting. 
The Committee welcomes suggestions on ways to increase the number of con- 
tributions to this annual survey, and to increase awareness of its customized com- 
puter search capabilities. 

Each literature reference or work in progress given below may contain the 
following, separated by semicolons: author(s); date; title; citation; and Indiana 
counties in which the work was performed. If any of these are absent, the con- 
tributor did not provide it. Counties are abbreviated by using their first five let- 
ters. The exception is Saint Joseph, which is abbreviated StJoe. 

An "ALL" in the county field indicates that the work applies to all 92 Indiana 
counties. A "MANY" indicates the work applies to many counties. In both cases 
up to five of the most important counties also may be included. 

34 



Biological Survey Committee Report 35 

Each citation begins with a letter which has one of the following meanings: 
P : Formal Publication (PIAS refers to Proceedings, Indiana Academy of 

Science) 
I : In Press in a formal publication 
T : Thesis 

: Other (semi) published work, e.g. park checklists, maps. 
E : Environmental Impact Statements 
W : Work in Progress 

Bennett, G. W. and E. S. Runstrom: 1980; Efficacy of fenthion, sodium bicarbon- 
ate, microencapsulated diazinon, and carbaryl against German cockroaches in 
urban housing; I: Journal of Economic Entomology; Mario. 

Bernhard, K. M. and G. W. Bennett; 1980; Ultra-low volume applications of 
synergized Pyrethrins for stored products pest control; I: J. Econ. Entomol. 

Brown, K. M. and A. Konopka; 1980; Interactions of phytoplankton and zooplank- 
ton in Crooked Lake; W: Dept of Interior grant; Noble Whitl. 

Brown, K. M.; 1980; Life history variation and competition in pond snails; W: 
NSF Grant; Allen Noble Whitl. 

Davies, W.; 1980; Seasonal variation and productivity of phytopsammon (interstial 
algae) of an artificial sandy beach; W: Purdue University; Whitl Noble. 

Deyrup, M.; 1980; Annoted list of Indiana Scolytidae; I: Great Lakes Entomolo- 
gist; Tippe Porte Clark Warre. 

Dineen, C. F.; 1979; Plankton and benthos of Spicer Lake; I: PIAS 89: 173-179; 
StJoe. 

Dineen, C.F.; 1980; Fishes of Spices Lake; I: PIAS 90; StJoe. 

Ferris, V. R., C. G. Goseco, and J. M. Ferris; 1980; Revisions of Oxydirus and 
Tarjanius n. gen. in Oxydiridae, Belondiroidea (Nematoda; Dorylaimida); and 
Oxydiroides in Prodorylaimidae, Dorylaimoidea; P: Purdue University 
Research Bull. 965:29 pp.; Riple. 

Goodman, J. D.; 1980; Cepedietta Kay, 1942 in the northern Ringneck Snake 
in Indiana; I: PIAS 90; Montg. 

Haddock, J. D.; 1980; Biological control of mealybugs by Crytolaemus sp. and 
other coccinelid predators; W: Purdue University-Fort Wayne; Allen. 

Hart, J. W. and R. D. Waltz; 1980; The Collembola of Indiana; ALL. 

Hendricks, D. R.; 1976; Checklist of woody plants native to the Whitewater 
Valley drainage Basin of Indiana and Ohio; 0: Hayes Regional Arboretum, 22 
pgs.; White Valle Wayne Fayet Frank Dearb. (10 counties) 

Hendricks, E. G.; 1977; Phenology of wildflowers at Hayes Regional Arboretum; 
0: Hayes Regional Arboretum, 48 pgs.; Wayne. 

Hendricks, D.R.; 1979; Tree S.A.V.E.R. (computer analysis of native tree species 
for homeowner planting); O: Hayes Regional Arboretum, 2 pgs; White Valle 
Wayne Fayet Frank Dearb. (10 counties) 

Hendricks, D. R. and Hendricks, E. G.; 1980; Flora of the Whitewater Valley 
Drainage Basin of Indiana & Ohio; W: Hayes Regional Arboretum; White Valle 
Wayne Fayet Frank Dearb. 

Iverson, J. B.; 1980; Demography of the Painted Turtle, Chrysemys picta; W; 
Kosci. 

Kozel, T. R.; G. W. Weddle, K. Welborn; 1980; A fish faunal survey of Posey 
County, Indiana; I: PIAS; Posey. 



36 Indiana Academy of Science 

Kruger, R. M. and R. R. Pinger; Larval Study of mosquitoes of Delaware county; 
W: Ball State; Delaw. 

Lawson, H. R. and L. Chandler; 1980; Neuroptera of Indiana: Checklist and 

key to the adults; I: In Press; ALL; Tippe Porte Lawre Warre. 
Lulla, K.; 1980; Detection of biomass changes in forest covers of Indiana using 

Landsat MSS data; P: Program Abstracts, 1980 ESA Meeting; Vigo. 
MacDonald, J. F.; 1980; Biology, recognition, medical importance and control of 

Indiana social wasps; P: Cooperative Extension Survey Bulletin E-91:23 pp; 

ALL. 

MacDonald, J. F., R. D. Akre: 1980; The German yellowjacket (Vespula ger- 

manica) problem in the United States (Hymenoptera; Vespidae); I: Entom. Soc. 

Amer., Bulletin 26 (4); Tippe. 
Marenchin, G., and D. M. Sever; 1981 Survey of the fishes of the St. Joseph 

River drainage in St. Joseph and Elkhart Counties, Indiana; I: PIAS 90; StJoe 

Elkha. 
Maxwell, R. H.; 1980; Indiana plant distribution records, Clark County; P: PIAS 

89:355; Clark. 
Maxwell, R. H.; 1980; Indiana plant distribution records; I: PIAS; Clark Floyd 

Harri Jeffe. 
Maxwell, R. H.; 1981; Checklist of vascular plants of the Indiana Army Ammunit- 
ion Plant, Clark County; W: Contribution of the Indiana University Southeast 

Herbarium; Clark. 
McCafferty, W. P.; 1979; Swarm-feeding by the damselfly Hetaerina americana 

(Odonata; Calopterygidae) on mayfly hatches; P: Aquatic Insects 1: 149-151; 

Marti. 
McCafferty, W. P. and M. C. Minno; 1979; The aquatic and semiaquatic Lepidop- 

tera of Indiana and adjacent areas; P: Great Lakes Entomologist 12: 179-187; 

Lake. Dekal Tippe Posey Porte. 
Meyer, R. W.; 1980; Insects and other arthropods of economic importance in 

Indiana during 1979; P: PIAS 89: 210-214; Harri Porte Warre Jacks. 
Miller, D. E.; 1980; Biology of a population of Ambystoma tremblayi; W: Ball 

State University — Dr. James C. List, Dept. of Biology; Delaw. 
Mould, E. D.; 1981; Nutritional status of Indiana squirrels; W: St. Mary's College, 

Eric Mould; StJoe Elkha LaPor. 
Parman, V. and M. C. Wilson; 1980; Interaction between the meadow spittlebug 

(Philaenus spumarius (Homoptera: Cerocopidae) and alfalfa (Medicago sativa); 

Warre. 
Richeson M. L., L. Herrman; 1980; Etiology of wilt in Helianthus annuus; W: 

Indiana University — Purdue University at Fort Wayne Biology Department; 

Allen. 
Riemenschneider, V.; 1980; Vascular plants of Sand Hill Nature Preserve, Pulaski 

County, Indiana; I: PIAS; Pulas. 
Riemenschneider, V.; 1980; Vascular plants of Barker Woods Nature Preserve, 

Laporte County, Indiana; W: Indiana University at South Bend; Lapor. 
Runstrom, E. S., G. W. Bennett, S. Ehrich, V. L. Anderson; 1980; Use of statis- 
tics in the comparative analysis of urban pesticide efficacy evaluations; I: J. 

Econ. Entomol.; Mario. 



Biological Survey Committee Report 37 

Shelton, M. D. and C. R. Edwards; 1980; Effects of weeds on the diversity and 
abundance of insects in soybeans; I: Environmental Entomology; Lawre. 

Sosa, 0. Jr.; 1980; Biotypes J and L of the Hessian fly discovered in an Indiana 
wheat field; I: Journal of Economic Entomology; Rando. 

Sparling, D. W.; 1980; Behavioral significance of plumage variability in wintering 
House Sparrows; W: Ball State University; Black. Delaw. 

Squters, E.; 1980; The effects of the season of fallowing on the development of 
first and second year old fields; W: E.R. Squiers, Taylor University; Grant. 

Torke, B.; 1980; Distribution of meiobenthic fauna in southern Lake Michigan; W: 
Biology Department, Ball State University; 

Walker, G. L. and R. E. Williams; 1980; A unique fly problem in an industrial 
bio-filter system; I: Journal of the Water Pollution Control; Tippe. 

Waltz, R. D.; 1980; The Caddisflies (Trichoptera) of Indiana; T: Purdue Univer- 
sity, Dr. W. P. McCafferty, Purdue University; ALL. 

Whitman, R.; 1980; Evaluation of Salt Creek (Indiana) for supporting and spawn- 
ing Coho Salmon; W: Indiana University Northwest, Biology Department; 
Porte. 

Williams, R. E. and J. G. Berry; 1980; CGA 72662 as a topical spray and feed 
additive for controlling houseflies breeding in chicken manure; I: Poultry 
Science; Carro. Tippe. 

Williams, R. E. and J. G. Berry; 1980; Control of northern fowl mite with perme- 
trin and fenvalerate, two synthetic pyrethroid compounds; I: Poultry Science; 
Tippe. 

Williams, R. E., T. L. McCain, and A. Taklahaimanot; 1980; Survey of pesti- 
cide usage by livestock producers in Indiana; 0: Research Bulletin; ALL. 

Williams, R. E., E. J. Westby, K. S. Hendrix, and R. P. Lamenager; 1980; Use of 
insecticide-impregnated ear tags for control of face flies and horn flies on 
pastured cattle; I: Journal of Animal Science; Duboi Grant. 

Williams, R. E. and E. J. Westly; 1980; The use of synthetic pyrethoids impre- 
gnated in cattle ear tags for control of face flies and hornflifes; I: Journal of 
Economic Entomology; Warre. 

Williams, G. E. and R. E. Williams; 1980; Survey of cattle grub populations in 
native Indiana cattle (Diptera: Oestridae); Publication; Lagra Porte. Washi 
Vande. 

Williams, W. T.; 1980; Environmental stress to forests & agroecosystems; I: 
Phytopathology; ALL. 



Necrology 
Fay Kenoyer Daily, Butler University 



Max William Gardner 

Lansing, Michigan Berkeley, California 

May 11, 1890 October 31, 1979 

Dr. Max W. Gardner died of a heart attack in Berkeley, California, October 
31, 1979. He was a botanist and Emeritus Professor of Plant Pathology at the 
University of California at Berkeley. 

He was interested in nature and Indian artifacts from early life, and studied 
wildlife around Lansing, Michigan, where he was born May 11, 1890. He kept an 
extensive diary relating his field studies; it was recently typed and preserved by 
his grandson, William Gardner Schottstaedt. The diary covers 1904 to 1911. He 
was particularly interested in birds and learned to identify them by song, flight 
patterns and behavior in addition to morphology. 

His early education was obtained in North Lansing schools. A B.S. degree in 
1912 and D.Sc. in 1950 were obtained at Michigan State University, and a M.S. 
degree in 1915 and Ph.D. in 1918 were bestowed by the University of Wisconsin. 
It was his service on the Pennsylvania Chestnut Blight Commission in 
Philadelphia that led F. D. Heald, with whom he worked, to encourage him to do 
graduate work with L. R. Jones at the University of Wisconsin. 

Dr. Gardner's first academic position was held at the University of Michigan, 
Ann Arbor, from 1917 to 1918. He was then Assistant Pathologist for the Bureau 
of Plant Industry, U. S. Department of Agriculture 1918 to 1919. In 1919, Dr. 
Gardner moved to Indiana. He was Botanist in the Experiment Station of Purdue 
University from 1919 to 1929 and was Chief of Botany from 1929 to 1932. While 
there, he initiated a graduate studies program. In 1932, Dr. Gardner moved to 
California. He became Professor of Plant Pathology and Plant Pathologist at the 
Experiment Station on the University of California, Berkeley and Davis. The ma- 
jor course given by him there was on diseases of truck crops. He became Emeritus 
Professor in 1957. After retirement, he continued building the reprint collection in 
the U. C. Plant Pathology Library, and collecting powdery mildews, many not 
previously described in the United States of America. 

About 180 publications were authored or co-authored by Dr. Gardner cover- 
ing various plant diseases and lightning injury to plants. He was associated in 
research with J. B. Kendrick, F. D. Heald, R. D. Studhalter, L. R. Jones, W. W. 
Gilbert, G. K. K. Link, H. S. Jackson, H. D. Brown, L. Greene, C. E. Baker, F. P. 
Cullinan, P. H. Brewer, H. R. Kraybill, L. C. Cochran, C. L. Burkholder, R. W. 
Sampson, H. E. White, E. B. Mains, J. J. Davis, C. M. Tompkins, 0. C. Whipple, C. 
M. Tucker, B. L. Richards, P. A. Ark, H. R. Thomas, J. T. Middleton, C. E. Yar- 
wood, A. E. Michelbacker, Ray F. Smith, P. G. Smith, E. B. Babcock, R. E. Smith, 
E. E. Wilson, R. D. Raabe, H. Kern, P. D. Caldis, T. Duafala, C. J. Krass, and W. 
N. Takahashi. Some of these were also Indiana Academy of Science members. 






Necrology 39 

Dr. Gardner joined the Indiana Academy of Science the year he came to Pur- 
due in 1919, and he was honored as Fellow in 1923. He gave a series of papers 
before the Academy at fall meetings from 1919 to 1924. He also co-authored a 
paper with J. B. Kendrick on a simple way to determine the thermal death point. 
He served on the Indiana Academy of Science Publications Committee from 1924 
to 1929 and the Anniversary Publicity Committee from 1931 to 1932. 

Dr. Gardner was active in other groups, too. He was on a committee in the 
Division of Biology and Agriculture of the National Research Council. He was also 
a life member of the American Phytopathological Society (v. pres. 1930, pres. 
1931, journal editor 1959 to 1964). He was a member of: Botanical Society of 
America, American Society of Naturalists, American Association for the Ad- 
vancement of Science, the Michigan Academy of Science, Gamma Alpha, and 
Sigma Xi. 

He and his wife, Margaret Briggs Gardner, have two children in the scientific 
profession: Mary Frances Schottstaedt, M. D., Associate Professor of Psychiatry, 
University of Texas Medical Branch at Galveston; and Murray Briggs Gardner, 
M. D. Professor of Pathology, University of Southern California School of 
Medicine. There are eight grandchildren. 

Dr. Gardner is listed in Indiana Scientists and American Men of Science. A 
very fine article about Dr. Gardner was written by C. E. Yarwood 
(Phytopathology 70(2): 79. 1980.), one of Dr. Gardner's first graduate students and 
close associate for 50 years. Dr. Gardner was a kind, considerate man and in Dr. 
Yarwood's words: "graciously shared his knowledge and inspired in his friends 
and family a love and respect for nature." 



40 Indiana Academy of Science 

Karl S(tone) Means 

Morristown, Indiana Indianapolis, Indiana 

January 2, 1892 July 18, 1980 

Dr. Karl S. Means was born in the small community of Morristown, Indiana, 
January 2, 1892. 

His college education began at Butler University where he received an A.B. 
degree in 1914. He then went to Indiana University for a M.A. degree in 1915 and 
received a Ph.D. degree in organic chemistry at Chicago University in 1924. 

Dr. Means first taught chemistry at Kokomo High School in 1916. Then he 
was at Butler University from 1917 to 1920; Chicago University 1920 to 1922; 
Michigan College of Mines from 1922 to 1926; Milligan College in Tennessee from 
1926 to 1928; and, back to Butler University from 1928 to 1957. He became full 
professor in chemistry in 1939. Dr. Means became Emeritus Professor at retire- 
ment. 

The chief research interests of Dr. Means were concerned with the electro 
deposition of antimony (Master's thesis) and the saponification of ethyl formate 
(Ph.D. thesis). At Butler, he was on the Men's Council, Student Affairs Commit- 
tee, special adviser for the Liberal Arts College, Pharmacy Adviser, adviser for 
the Butler University Independent Association (student society) and adviser for 
Alpha Chi Omega. He gave significant service in his church also, as chairman of 
the Board of Elders, Chairman of the Education Committee, member of the Board 
of Directors, as well as the Marion County Association of Christian Schools. 

Dr. Means joined the Indiana Academy of Science in 1928 and became a 
Fellow in 1940. He was an Emeritus Member at death. He gave a paper at the fall 
meeting of the Academy in 1940 on the relation of valence and nomenclature to 
formulas for negative ions of ternary inorganic compounds. He was Division 
Chairman of the Chemistry Division that year. From 1942 to 1951, he was on the 
Auditing Committee and served as Chairman from 1947 to 1951 when he was also 
on the Executive Committee. He served on the Program Committee in 1944. 

Dr. Means belonged to several other societies: the American Chemical Society, 
Alpha Chi Sigma, and Indiana Chemical Society, and was honored by election to 
Phi Kappa Phi and Sigma Xi. 

Dr. Means died July 18, 1980, at Indianapolis, Indiana, at 88 years of age. He 
had served long and well as teacher and counselor at Butler University and in the 
many duties of his religious and civic activities. He was a quiet, kind and depend- 
able man. 



INDIANA ACADEMY OF SCIENCE 
NEW MEMBERS -1980 

Adams, Dr. Preston, Dept. of Botany & Bacteriology, DePauw University, Green- 
castle, IN 46135 
Aldrich, Mr. James R., 612 State Office Bldg., Indianapolis, IN 46204 
Alexander, Mr. Lee H., 208 E. 6th St., Muncie, IN 47302 
Arnold, Dr. Gerald B., Dept. of Physics, Univ. of Notre Dame, Notre Dame, IN 

46556 
Bartmess, Dr. John E., Dept. of Chemistry, Indiana University, Bloomington, IN 

47405 
Beeler, Miss Cynthia L., 4356 Buchanan St., Gary, IN 46408 
Black, Ms. Charlene, 2001 Roosevelt St., Gary, IN 46404 
Bowden, Dr. Warren W„ 5500 Wabash Ave., Rose-Hulman Inst, of Technology, 

Terre Haute, IN 47803 
Braun, Mr. Edward R., Tri-Lakes Fisheries Station, R.R. 4, Columbia City, IN 

46725 
Bray, Dr. Malcolm D., R.R. 1, Box 710, Noblesville, IN 46060 
Bright, Mr. Greg R., Ind. State Board of Health, 1330 W. Michigan, Indianapolis, 

IN 46206 
Brunson, Dr. Kenneth W., 3400 Broadway, Ind. Univ. School of Medicine, Gary, 

IN 46408 
Burrows, Mr. Stephen D., 401 Sherwood Forest, Galveston, IN 46932 
Bush, Dr. Kenneth H., 1130 N. Salisbury St., W. Lafayette, IN 47906 
Butler, Dr. Frank, Dean, IU East, Richmond, IN 47374 
Cantrell, Mr. Buddy, Dept. of Biology, Univ. of Notre Dame, Notre Dame, IN 

46556 
Carpenter, Dr. Stephen R., Dept. of Biology, Univ. of Notre Dame, Notre Dame, 

IN 46556 
Chesak, Dr. David D., 420 S. Weston St., Rensselaer, IN 47978 
Clemens, Dr. Donald G., Dept. of Chemistry, Goshen College, Goshen, IN 46526 
Couts, Mr. Paul J., 715 N. Earl Ave., Lafayette, IN 47904 
Datta, Mr. Bithin, 745 Grad. Housing East, W. Lafayette, IN 47906 
Dilling, Dr. Richard A., Grace College, Winona Lake, IN 46590 
Dunn, Mrs. Vivie E., Ind. State Board of Health, 215 Monroe, Newburgh, IN 

47630 
Ellis, Mr. Mark G., P.O. Box 74, Albany, IN 47320 
Farster, Ms. Jill Ann, 324 W. 41st Ave., Gary, IN 46408 
Ferson, Mr. Scott David, Dept. of Ecology & Evolution, SUNY at Stony Brook, 

Stony Brook, NY 11794 
Fogle, Dr. Thomas, Dept. of Biology, St. Mary's College, Notre Dame, IN 46556 
Frankowski, Ms. Sheryl, 4870 Jefferson St., Gary, IN 46408 
Franz, Dr. Frank A., Bryan Hall 109, Indiana University, Bloomington, IN 47405 
Garner, Mr. John, Cloverdale High School, Cloverdale, IN 46120 
Gillespie, Miss Sharon L., 6818 Maryland Ave., Hammond, IN 46323 
Green, Dr. John G., Box 35, Huntington College, Huntington, IN 46750 
Greengold, Mr. Gerald, Dept. of Geosciences, Purdue University, W. Lafayette, 

IN 47906 

41 



42 Indiana Academy of Science 

Greenlee, Ms. Judith, Dept. of Biology, Univ. of Notre Dame, Notre Dame, IN 

46556 
Grimstad, Dr. Paul R., Dept. of Biology, Univ. of Notre Dame, Notre Dame, IN 

46556 
Gulino, Ms. Terri, 750 West Hampton Dr., Indianapolis, IN 46208 
Hailer, Mr. Joseph G., 611 N. Walnut Grove, Bloomington, IN 47405 
Hall, Mr. Bradley K., 509 Travers Circle, Apt. D, Mishawaka, IN 46544 
Hamrick, Mrs. Linda, Canterbury School, 5601 Covington, Rd., Ft. Wayne, IN 

46819 
Hasenmueller, Mr. Walter A., Ind. Geological Survey, 611 N. Walnut Grove, 

Bloomington, IN 47405 
Haughs, Mr. R. Douglas, 6900 Bloomfield Dr. E., Indianapolis, IN 46259 
Hawkins, Dr. M. Jeanne, 636 Linwood #218, Buffalo, NY 14209 
Hawthorne, Dr. Robert M., Jr., Purdue University-North Central Campus, West- 

ville, IN 46391 
Hedge, Mr. Cloyce L., 612 State Office Bldg., Indianapolis, IN 46204 
Herrman, Dr. Larry, Dept. of Biology, IU-PU at Ft. Wayne, Ft. Wayne, IN 46805 
Hively, Dr. Ray M., Dept. of Physics, Earlham College, Richmond, IN 47374 
Hoffman, Mrs. Sarah, R.R. 3, Box 41, Rochester, IN 46975 
Holdiman, Ms. Jeannine, 750 W. Hampton Dr., Indianapolis, IN 46208 
Holland, Dr. James Philip, 4125 South Gran Haven Dr., Bloomington, IN 47401 
Hosek, Mr. Henry, Jr., Purdue University-Calumet Campus, Hammond, IN 46323 
Howes, Dr. Ruth, Dept. of Physics & Astronomy, Ball State University, Muncie, 

IN 47306 
Hudson, Mr. W. Donald, Jr., 115 S. Bryan St., Bloomington, IN 47401 
Huffman, Dr. John C, Molecular Structure Center, Indiana University, Blooming- 
ton, IN 47405 
Johnston, David K., Dept. of Geology, Ball State University, Muncie, IN 47306 
Jones, Dr. Barry Nelson, Roche Inst, of Molecular Biology, Nutley, NJ 07110 
Kaellner, Mr. Jeff W., Elliott Hall, Ball State University, Muncie, IN 47306 
Kendra, Mr. Albert, Dept. of Biology, Ball State University, Muncie, IN 47306 
Kirkpatrick, Dr. Ralph, Dept. of Biology, Ball State University, Muncie, IN 

47306 
Kowalski, Mr. Michael P., Dept. of Biology, Indiana University, Bloomington, IN 

47405 
Kyker, Dr. G. C, Jr., Dept. of Physics, Rose-Hulman Inst, of Technology, Terre 

Haute, IN 47803 
Lindmark, Dr. Alan F., 3503 Coventry Circle, Valparaiso, IN 46383 
Litton, Dr. James R., Jr., Dept. of Biology, St. Mary's College, Notre Dame, IN 

46556 
Longenecker, Mr. Nevin, 1510 Renfrew Dr., South Bend, IN 46614 
Lovell, Dr. C. W., School of Civil Engineering, Purdue University, W. Lafayette, 

IN 47906 
Loyd, Mr. K. Mark, Dept. of Natural Resources, Ball State University, Muncie, 

IN 47306 



New Members -1980 43 

Lyon, Dr. Ed, Dept. of Geography/Geology, Ball State University, Muncie, IN 

47306 
McClain, Mr. William F., Illinois Dept. of Conservation, 605 Stratton Building, 

Springfield, IL 62706 
McDonald, Mr. Eric, 514V2 West High St., Elkhart, IN 46514 
McKinney, Dr. Paul C, Wabash College, Crawfordsville, IN 47933 
McNitt, Mr. Timothy J., 2803 Parnell Ave., Ft. Wayne, IN 46805 
McTague, Mr. Michael, 818 W. Riverside Ave., Apt. E4, Muncie, IN 47303 
Magenheimer, Ms. Cindy, 750 West Hampton, Indianapolis, IN 46208 
Mallett, Dr. Gordon E., Eli Lilly & Co., 307 E. McCarty St., Indianapolis, IN 

46285 
Martin, Mr. Patrick P., R.R. 16, Box 393, Brazil, IN 47834 
Matney, Ms. Elizabeth Ann, 406 Western Dr., Bloomington, IN 47401 
Mayes, Dr. Richard Alan, Dept. of Botany & Bacteriology, DePauw University, 

Greencastle, IN 46135 
Midriff, Ms. Karen R., 2103 W. Washington, #4, Muncie, IN 47303 
Miller, Mrs. Kimberly, I6OOV2 West Jackson St., Muncie, IN 47303 
Mitchell, Dr. George R., Purdue University-Calumet Campus, Hammond, IN 

46323 
Mosher, Mr. Dan, 5707 River Rd., Muncie, IN 47304 

Mould, Dr. Eric, Dept. of Biology, St. Mary's College, Notre Dame, IN 46556 
NlCKELL, Mr. Allan K., U. S. Soil Conservation Service, 1417 Bear St., Madison, 

IN 47250 
O'Loughlin, Dr. Carol L., 1201 East 38th St., Indianapolis, IN 46205 
Orr, Mr. John W., Ind. State Board of Health, 1330 W. Michigan St., Indiana- 
polis, IN 46206 
Ortman, Dr. Eldon, 3615 Capilano, W. Lafayette, IN 47907 
Provost, Dr. Paul-Jean, Dept. of Anthropology, IU-PU at Ft. Wayne, Ft. Wayne, 

IN 46805 
Radcliffe, Mr. Michael J., Redbud Hill, Apt. 603, Bloomington, IN 47401 
Raubenheimer, Miss Kim J., 3535 West 14th St., Indianapolis, IN 46222 
Reed, Dr. David K., USDA-SEA-AR, P.O. Box 944, Vincennes, IN 47591 
Rosen, Dr. S. P., Dept. of Physics, Purdue University, W. Lafayette, IN 47906 
Rumbaugh, Max E., Jr., 1125 Brookside Ave., P.O. Box 80-B, Indianapolis, IN 

46206 
Schroeder, Dr. Dolores M., 3518 Tudor Lane, Bloomington, IN 47401 
Schwarzw alder, Mr. Robert, Jr., Dept. of Biology, Indiana University, Blooming- 
ton, IN 47405 
Shaffer, Mr. Nelson R., 611 N. Walnut Grove, Indiana Geological Survey, Bloom- 
ington, IN 47405 
Shipman, Mr. Stuart T., R.R. 4, Columbia City, IN 46725 
Siminski, Miss Cynthia, 6643 Maryland Ave., Hammond, IN 46323 
Sinclair, Mr. H. Raymond, Jr., 410 Montrose Court, Indianapolis, IN 46234 
Sousa, Dr. Lynn R., Dept. of Chemistry, Ball State University, Muncie, IN 47306 
Speece, Dr. Susan P., Decker Hall, Anderson College, Anderson, IN 46011 



44 Indiana Academy of Science 

Stabler, Dr. Timothy A., Dept. of Biology, IU Northwest, Gary, IN 46408 

Stewart, Mr. Michael James, 2010 South 6th, Terre Haute, IN 47802 

Stewart, William and Kathy, 1709 East Madison, South Bend, IN 46617 

Sweeney, Mr. Vernon D., 702 West 7th, Bloomington, IN 47401 

Taylor, Dr. James I., Box G, Univ. of Notre Dame, Notre Dame, IN 46556 

Thomas, dr. Gerald P., Dept. of Physics, Ball State University, Muncie, IN 47306 

Troxel, Ms. Karen S., 1122 South 21st St., Lafayette, IN 47905 

Vandenberge, Dr. James C, 10030 Fifth St., Highland, IN 46322 

Vaughan, Mr. Martin A., Dept. of Life Sciences, Indiana State University, Terre 

Haute, IN 47809 
Vaughn, Mr. Danny M., 812 North 7th St., Apt. B, Terre Haute, IN 47809 
Vogelmann, Mr. James, Dept. of Biology, Indiana University, Bloomington, IN 

47405 
Vyas, Mr. Dilip V., 215 North College Ave., Muncie, IN 47306 
Wagner, Ms. Wendy, ADPI House, Hanover College, Hanover, IN 47243 
Wallace, Mr. Howard A., P.O. Box 653, South Bend, IN 46624 
Weaver, Dr. Connie, Dept. of Foods & Nutrition, Purdue University, W. Lafay- 
ette, IN 47906 
Wenstrup, Mr. Jeffrey, 256C Myers Hall, Indiana University, Bloomington, IN 

47405 
Wesonga, Mr. Sam, 520 North Grant, #9, Bloomington, IN 47401 
Wright, Mr. Joe, Div. of Curriculum, Rm. 229, State House, Indianapolis, IN 

46204 
Yovits, Dr. Marshall C, 9016 Dewberry Court, Indianapolis, IN 46260 
Zimmer, Mr. Lester L., R.R. 1, Box 155, Nashville, IN 47448 
c/o Linda Hamrick Canterbury School Science Club, 5601 Covington Rd., Ft. 

Wayne, IN 46819 
c/o Larry D. Bennett, Mitchell Jr. High School Science Club, Hancock Ave., 

Mitchell, IN 47446 



ADDRESSES AND CONTRIBUTED PAPERS 



SCIENTIFIC RESEARCH AND ECONOMIC INDICATORS 

Robert E. Henderson 
Indianapolis Center for Advanced Research 

When C. P. Snow wrote his treatise on Two Worlds, he emphasized the dif- 
ferent world views of scientists and non-scientists (11). His viewpoint was empha- 
sized by the question, "How often is the second law of thermodynamics discussed 
at cocktail parties?" I have tried for the past ten years to do just that, but with 
limited success. Now Jeremy Rifkin has written a book, Entropy (9), based on the 
penetrating analysis by Georgescu-Roegen of economics and the second law (5). 

As we enter the eighties, it is necessary that all the tools at our command be 
brought to bear on social problems that we have come to call economic. I believe 
that C. P. Snow's Two Worlds in this matter relate to disinterest and self-interest 
on the part of scientists, who I believe have the necessary knowledge for purposes 
of solving these problems. On the other hand, social scientists and economists are 
making proposals such as enhancing productivity through basic research and 
other scientific activities, misleading us because of their lack of experience and 
understanding of current scientific techniques, culture, and organization. 

My comments are based on involvement in the American science/research 
arena for the past 30 years. This involves 20 years as a scientist, engineer, and ad- 
ministrator with an aerospace division of a large American automobile manufac- 
turer. Those twenty years were sandwiched between exposure to academic 
research in the early 50s as a graduate student; and, to academic and non-profit 
research in the late 70s as an administrator. My approach to administration is 
oriented toward motivating individuals to develop and make their scientific 
talents available by providing the appropriate instrumentation and equipment, 
clear-cut goals, and a demanding, but intellectual, environment. This has led me to 
believe that the two worlds gap is widening— not closing — and that we, as scien- 
tists, areJargely responsible. I am also aware that my comments may be deemed 
unsophisticated by economists, but risk that criticism in order to aid in developing 
the forum for communication. 

Let us review, briefly, the economic indicators related to our social problems. 
First, of course, is the question of the depressed value of currencies, in particular, 
the American dollar. Figure 1 is a plot showing that the cost of industrial 
materials has grown more rapidly than consumer prices since 1967. The latter 
have increased by a factor of 2V2 in that time period. At the same time, the 
balance of trade for the U.S. has been negative. Figure 2 demonstrates that oil 
shocks have modulated a basic trend showing a lack of competitiveness in the 
marketplace on the part of American products (8). For instance, in the days follow- 
ing the first oil boycott, OPEC countries used their surplus to import American 
products, thus, producing a surplus. This market was quickly saturated and the 
downward trend continued. We now are experiencing a second wave of that same 
effect which, unfortunately, will apparently not reach zero this time. 

An economic term called productivity has been identified as a significant 

45 



46 



Indiana Academy of Science 




1967 



Year 

Figure 1. Inflation 



10 



Oil -Embargo 





(0 

iS 

o " 10 

Q 



^ 



Period 1 



Slowly weakening technology 
because of misplaced 
priorities 



I I I I I I 

73 75 \ 77 79 



Period 2 



CO 

c 
o 



CO 



-20 



OPEC 
spending of 
new petro 
dollars 



Period 3 



30 




"40 



Year 

Figure 2. U. S. Trade Balance— All Commodities 



Presidential Address 



47 



Index 


: 1967=100 








230- 






• — • United States 








210- 






* — * France 






+ 


190- 






■ — ■■ West Germany 
♦ — + Japan 




/ 


/"X/ 


i 170 1 

_ 150^ 

<D 

| 130- 
? 110- 






— ° United Kingdom 
x — x Canada 


♦ — 






a> 

; go- 














f 70 i 




I^jN 




° 50- 


,| 


_____+ — 






















30- 














10- 














19 


60 


i 

62 


i I i i i i i 
64 66 68 


1 1 

70 


\ 

72 


i i i i i i 
74 76 78 



Year 

Figure 3. Productivity Change in Manufacturing Industries By Selected Coun- 
tries, 1969-1977 

culprit. Figure 3 and Table 1, however, show that although U. S. productivity is 
increasing more slowly than that of other countries, it is the highest in the world 
and is still increasing (3,2). 

Further, it has been argued that lack of research and development is the 
basis for lowered productivity; and, therefore, a root cause of economic problems. 

Table 1. Real Gross Domestic Product Per Employed Civilian, for Selected 
Countries Compared With the United States: 1960-77 

[Index. United States = 100] 





United 




West 




United 




Year 


States 


France 


Germany 


Japan 


Kingdom 


Canada 


1960 


100 


55.4 


52.4 


24.7 


51.1 


86.6 


1961 


100 


57.0 


53.1 


27.2 


49.8 


85.6 


1962 


100 


58.0 


53.1 


27.6 


47.9 


85.6 


1963 


100 


59.0 


53.2 


29.6 


48.5 


86.2 


1964 


100 


60.0 


55.2 


32.1 


49.1 


86.0 


1965 


100 


60.8 


56.2 


32.2 


48.2 


85.6 


1966 


100 


61.2 


58.1 


33.4 


47.4 


83.5 


1967 


100 


63.4 


57.3 


36.8 


49.0 


83.4 


1968 


100 


64.2 


59.5 


40.0 


49.7 


84.6 


1969 


100 


67.6 


63.2 


43.9. 


50.4 


86.2 


1970 


100 


71.4 


67.0 


48.7 


52.6 


88.6 


1971 


100 


72.9 


67.0 


48.7 


52.6 


88.6 


1972 


100 


74.8 


68.5 


53.9 


53.6 


90.7 


1973 


100 


76.4 


70.2 


56.5 


54.8 


90.8 


1974 


100 


80.0 


74.3 


58.0 


56.0 


93.0 


1975 


100 


81.2 


74.7 


59.5 


55.4 


91.9 


1976 


100 


83.1 


77.7 


60.8 


55.6 


92.2 


1977 (prel.l 


100 


84.7 


79.1 


62.2 


55.1 


91.6 



SOURCE: Department of Labor, Bureau of Labor Statistics 



48 



Indiana Academy of Science 



Percent 
3.8 



o 

Q. 

75 

c 
o 

+■> 



(0 
(0 
O 



1.8 



14 ,' 



O 1.0 



0.6 



0.2 




/ 



/ /^ Germany .•**' 
V. ...... ,...--' s Japan 



ii' i i i i i i I i 



1961 63 65 67 69 71 

Years 



73 75 77 78 



Source: Science Indicators -1978 



Figure 4. National Expenditures for Performance of R&D as a Percent of GNP 

by Country, 1961-78 



Presidential Address 49 

Figure 4 demonstrates that, on the contrary, U. S. R&D efforts are supported not 
only at a higher dollar level than any other country in the West, but also at a 
higher level of percent Gross National Product (10). I would add that the recent 
Nobel awards emphasize the fact that the U. S. is the world leader in scientific 
research. In the magazine, Science, a speech by Lewis Branscomb of IBM Cor- 
poration was recently condensed as a relevant editorial. He pointed out that we 
are still world leaders in science, our engineering talents are still tops, and the 
quality of our technology is still the highest in the world. He summarized by say- 
ing, "The picture of American science and technology today is one of great 
strengths yet deep doubts; of strong foundations and timid commitment; of critical 
importance to the economy and uncertain political priority. If indeed our domestic 
and our foreign trade performance are poor, is lagging technology the symptom or 
the cause? And if technology lags, is this because the steam has gone out of our 
science? Or because of a failure of economic policy and industrial will?" 1 

Although there are many versions of cause and effect for these ills, they 
generally take the form of decrying lack of support for a given ingredient. For in- 
stance, certain manufacturers call for trade restrictions, the construction industry 
for lower interest rates, and economists discuss taxes and tax incentives at great 
length. In scientific circles, the connection is made between R&D support, innova- 
tion, and productivity. Thus, spokesmen for science are calling for ever greater 
support in more or less traditional form as a major component in improving 
American productivity and, thereby, combating inflation. 

Another major source of inflation is clearly the increase in the price of oil; 
and, to a lesser extent, other strategic materials. These increased prices feed 
through the economy and increase the cost; and, therefore, the price of all goods, 
and especially food stuffs, since American farming techniques have become highly 
energy intensive. Once again the proposals made to solve this aspect of inflation 
are generally related to the needs of the petitioner to a greater extent than the 
solving of the problem. For instance, consumer groups call for more price controls, 
oil companies for lifting of environmental restrictions and the opening of more 
regions to exploration and drilling, and economists once again discuss tax and sub- 
sidy approaches. Engineers and scientists recognized at an early stage the need 
for their expertise in meeting this challenge. However, the outcome has been a 
wealth of self-serving proposals generally emphasizing that support of R&D will 
reap energy benefits in the future. A good example is the politicization of fusion 
research. This interesting, but costly, scheme is not relevant to the liquid fuel 
crunch of the eighties, but its public relations announcements are timed to concur 
with congressional budget hearings. 

In summary, the consensus scenario is that productivity decline and foreign 
energy monopolies, together with normal inflationary pressures (wage demands), 
have caused a serious social problem called inflation (3). Our federal government 
is responding to pleas from various constituencies to provide support to those 
special interest groups in order to solve the problem. In particular, the science 
establishment is calling for greater and greater support of R&D in order to 
enhance productivity and replace foreign oil. 

I believe there is a serious deficiency in this scenario of increasing R&D 
budgets (already high) as a response to inflation, unless we make major changes in 
the way we do our research. It seems to me that we are not managing the R&D 



Lewis Branscomb, "Needed: Conviction to Match Our Science," Science, August 1980, Vol. 309, No. 4457, 
p. 641 (excerpted from a commencement address at Polytechnic Institute of New York, May 29, 1980). 



50 Indiana Academy of Science 

establishment in a way that is beneficial to the nation and that this ineffectiveness 
has three causes: 

1. Insufficient knowledge on the part of decision makers. The combination 
of economists, industry leaders, governmental bureaucrats and elder scientific 
statesmen lack knowledge of modern hardware technologies. The extremely rapid 
growth and complexity of such fields as microelectronics makes anything but 
direct experience with laboratory work or design cause rapid obsolescence in 
scientific knowledge. For example, some governmental officials do not have a 
realistic grasp of what research is, how it is carried out, what it can do, and what 
its limitations are. The more complex the methodology, the more mysterious it 
seems to those who are not privy. This leads to an understandable frustration in 
that these officials have an unrealistic expectation of solid answers being made 
available to major problems fairly quickly. 

2. Conflicts of interest. All parties related to these problems perceive of 
them in a way that a greater need for their own expertise is required. For scien- 
tists, this has all too often amounted to requests for continuation of conventional 
research. Certainly, for example, there are different roles that researchers and 
policy makers must play. Many, if not most, policy issues have important political 
components that policy makers must weigh — one of our jobs as researchers, then, 
is to insure that we take into account the non-technical aspects of our research 
projects as well as the technical ones. More about this later. 

3. Time lag phenomena. The symptoms of economic distress relate to 
causes which predate them by many years, in most cases. Analysts are not clear 
on either the nature of causes or the appropriate time lags. A good example of 
long-term innovation is the Cable TV industry. Work began on the research and 
development of this method of bringing the TV signal into the TV set as far back 
as the early 1930s. Forty-four years later we are finally seeing the rapid rise of 
marketing and installing this system for community use. Many patents and much 
research funding failed to bear fruit for many, many years; but persistent effort to 
bring this innovation to commercialization appears finally to be a reality. 

I do not have a well-prepared solution to these many problems. There is little 
time in this talk to develop song and verse to rebut the conventional economics, 
not to mention support a different scenario. I would, however, like to express a 
possible viewpoint based on my own experience — one which is markedly different 
from those of present decision makers. Possibly this viewpoint could be analyzed 
by others for its value and added to the mix of propositions for problem solution. 

The scenario, as I see it, is as follows: 

1. The Productivity Index is an economist invention which is nearly mean- 
ingless. Its slowing growth rate probably relates to the steady increase in the ser- 
vice industry sector. 

2. In the United States, innovation normally comes through smaller in- 
dustries. The venture capital drought in the mid-1970s made this impossible. A 
five-year time lag makes this problem now evident. 

3. Our energy problem is a result of consistent government cheap energy 
policies. This is a case of legislating science. The time lag involved here is of the 
order of ten years. 

4. Our trade imbalance is due to the innovation slump mentioned previously 
with a major downward revision due to energy policy. 



Presidential Address 51 

5. Inflation comes about because of increased dependence on service in- 
dustries for growth in U. S. and the trade balance deficit. 

6. The administration of R&D nationally has been poor and has been coor- 
dinated with the cheap energy policy. A major problem has been the type of 
research conducted, not the amount of research. 

Before proceeding with further comments regarding my views of the root 
causes of our economic trauma, a short history of U. S. research and development 
taxonomy is in order. 

Since World War II, research and development have been defined as follows 

(1): 

Basic Research. Basic research is that research which is directed toward increase of knowledge in 
science. The primary aim of basic research is a fuller knowledge or understanding of the subject under 
study, rather than any practical application thereof. 

Applied Research. Applied research is that effort which (a) normally follows basic research, but may 
not be severable from the related basic research, (b) attempts to determine and exploit the potential of 
scientific discoveries or improvements in technology, materials, processes, methods, devices, or 
techniques; and, (c) attempts to advance the state of the art. Applied research does not include efforts 
whose principal aim is design, development, or test of specific items or services to be considered for 
sale; these efforts are within the definition of the term development, defined below. 

Development. Development is the systematic use, under whatever name, of scientific and technical 
knowledge in the design, development, test, or evaluation of a potential new product or service (or of 
an improvement in an existing product or service) for the purpose of meeting specific performance re- 
quirements or objectives. Development includes the functions of design engineering, prototyping, and 
engineering testing. Development excludes subcontracted technical effort, which is for the sole pur- 
pose of developing an additional source for an existing product. 

Systems Analysis. Systems and other concept formulation studies are analyses and study efforts either 
related to specific IR&D efforts or directed toward the identification of desirable new systems, 
equipments or components, or desirable modifications and improvements to existing systems, 
equipments, or components. 

Of course, slightly different versions of these definitions abound. 

Many new definitions and experiments in research and development 
methodology have been undertaken in recent years. In general, the purpose of 
these studies has centered around attempts to increase productivity as measured 
by such parameters as number of publications, patent activity, product innova- 
tion, and societal impact. 

One of the major problems confronting research administrators, in this age of 
high costs, is that of increasing productivity. Measuring productivity is difficult at 
best; but, one way of doing it in the research field is to use the relationship be- 
tween societal benefit from research to its cost. 

Numerous problems are inherent in this methodology. The most important is 
that of the time constant between time at which research is conducted and time at 
which benefit is derived. Research managers have historically approached this by 
either selecting short-range projects or by breaking longer-range projects into 
definable shorter-range milestones. There is some question whether the first ap- 
proach is research at all, while most research scientists view the latter as an un- 
necessary activity at best and more often substitute the intermediate goals for 
the social benefit. This, in turn, tends to further the already growing division be- 
tween science and society. Good examples of the failure of this approach are the 
figures of merit developed by thermoelectricity groups in the 50s and by fusion 
groups in the 60s. The figures of merit were broken into material properties 
which set off research projects intended to produce those specific properties. 
Research efforts soon diffused and large expenditures led to no social benefits. 



52 



Indiana Academy of Science 



The development of a refined approach to improving R&D performance, by 
linking research more directly to a process, we have come to call innovation. In- 
novation is defined as that process by which a new idea is successfully translated 
into economic impact within our society. It is the introduction, sale or use of a new 
technology — whether product, process, or system, in the consumer or industrial 
marketplace. Thus, innovation encompasses the entire spectrum of research, 
development, invention, finance, marketing, production, management and sales. 

Many studies have been conducted which link innovation in technology to 
small companies in the private sector. A recent study of this type made by John 
Gilman of Allied Chemical resulted in the graph in Figure 5, which plots inventi- 



? 1000 
o 



CO 
0) 



100 



■♦-» 



10 



Regression Line 




0.1 



10 



100 



Sales (10 9 $/yr) 

Figure 5. Inventivity in Patents/Sales 



Presidential Address 53 

vity in patents/sales versus sales for American corporations (7). Note the inverse 
relationship. Statistically, according to Gilman, the inverse dependence is 
relatively high with a coefficient of 0.79. In Table 2, venture capital developed in 
the U. S. for high technology firms as a function of the years through the 70s, 
demonstrates clearly that we have starved just those companies which can best 
speak to the question of innovation (4). Although recent increases in this type of 
investment are occurring, the time lag between company formation and commer- 
cial significance is such that no effects have been felt on the economy. 

Table 2. Venture Technology Public Offerings 1969-77 





Sma 


1 Technic- 


il F 


rms 


Number of 




Tot 


al Dollar Amount 


Offerings 


($ millions) 




($ millior 


s) 






1969 




1367 






698 


1970 




375 






198 


1971 




551 






248 


1972 




896 






409 


1973 




158 






69 


1974 




16 






9 


1975 




16 






4 


1976 




145 






29 


1977 




118 






30 



Source: MIT Development Foundation 

The explanation for this cause and effect has been described by George 
Gilder as nothing more than the battle between the old and the new, or the past 
and future (6). Government and industry have established factories, offices, and 
bureaucracies which will seem worthless in the face of new technologies. He 
quotes the oft quoted Joseph Schumpeter as saying, "creative destruction is the 
essential fact of capitalism." As a result, large industry and government tend to 
want to plan while new companies go ahead and implement. This is noticeable in 
the energy program. While the government spends millions on developing an 
energy plan, small companies spend thousands on building and selling solar collec- 
tors. 

The Indianapolis Center for Advanced Research, for the past seven years, 
has been conducting a living experiment in combining the various elements of 
R&D together with innovation and discovery to the end that the time lag between 
discovery and commercialization can be reduced. The results would thus be an im- 
provement in U. S. competitive capability through R&D. The results have been 
encouraging. The characteristics of this type of research are fourfold: 

1. Novelty and discovery. 

2. Mission orientation. For ICFAR this means efforts that foster either 

a. the improvement of the quality of life (cultural); 

b. the development of new products, enterprises, processes, and ser- 
vices which strengthen the free enterprise system (commercial); 

or, 

c. the development of nationally-recognized scholarly research in the 
life and physical sciences (academic). 

3. The time lag between initiation of research and implementation of the 
results due to social and other non-technical barriers is taken into account in 
research planning. 



54 Indiana Academy of Science 

4. A research style is employed which embodies special approaches to sur- 
mount social, economic, political, and other non-technical barriers to the applica- 
tion of results. 

This concept of advanced research, using novelty, creativity, mission orienta- 
tion, and time dependent planning, emphasizes the development of style. It is, 
therefore, appropriate to quote an acknowledged leader of research from another 
era, J. Robert Oppenheimer. He said: "It is style which complements affirmation 
with limitation and with humility; it is style which makes it possible to act effec- 
tively, but not absolutely; it is style which enables us to find harmony between the 
pursuit of ends essential to us, and the regard for the views, the sensibilities, the 
aspirations of others; it is style which is the deference that action pays to uncer- 
tainty; it is above all style through which power defers to reason." 2 

It is certain that we have had enormous development and diffusion of 
knowledge in the last three decades. This knowledge now transcends that to 
which we had previously become accustomed. It includes fantastic capabilities on 
the part of technicians, engineers, and scientists. The potential for accomplish- 
ment of objectives has been increased by orders of magnitude. We have come so 
far that the potential for accomplishment, for practical purposes, can be con- 
sidered limitless. This basic argument means that the important decisions to be 
made in research and development at the present time must deal with deciding 
what we wish to accomplish. What are the questions that need to be answered? 
What kinds of problems are those that we must address? These are not simple 
questions, because we have many, many different problems reported by different 
groups. Our politics at the present time, to a great extent, are single issue politics. 
Nevertheless, the question is more one of selection of what it is we want to solve 
and to accomplish, than one of stocking our shelves with a great deal more infor- 
mation. A second type of question takes the following form: Define the economic, 
social, and cultural barriers to the application of the new knowledge generated in 
a given field and how can the research be best structured to surmount them. 
Thus, my view of the administration of research and development programs today 
is to determine what new accomplishments we must achieve and how shall we 
overcome the economic and cultural barriers to the application of the results of 
the necessary research. 

Reviewing once again. . .With the enormous amount of knowledge and tech- 
nique that we have available at the present time, it is my contention that it is a 
question of selecting objectives and how to treat the environment surrounding the 
science in order to bring about the application of the results of research. 

Possibly a general example will be of help. We decry the depletion of our 
resources — everything from fossil fuels to critical metals such as cobalt are 
rumored to be, or in fact are, in short supply for various reasons. There are many 
approaches to the solution to these problems. First, recycling is an obvious ap- 
proach to regaining major amounts of natural resources, both in fossil fuels and 
for critical metals. A second is underwater mining. The sea has available enor- 
mous resources, including fossil fuels. The nation is undertaking some programs 
now in the Baltimore Canyon on the east coast. Major resources of natural gas lie 
under the Gulf of Mexico. The decision to go ahead with respect to programs of 
this sort is not a question of technical know-how or more knowledge. It is a ques- 
tion of resolve to overcome the associated problems and a willingness to recognize 



Trom Dr. Laurence J. Peter. Peter's Quotations-Ideas of Our Time. William Morrow and Company, Inc. 
1977. 



Presidential Address 55 

them as problems. I might point out that there is a third resource in space, which 
is essentially infinite in scope. This is a much more expensive route, but one 
worthy of longer-range considerations. 

Solutions such as these face economic and social barriers. These must be 
recognized and overcome by putting scientists and engineers to work in an ad- 
vanced research modality. We have not yet, of course, done that. The problems 
must be well defined and the barriers to the application of the results of the 
research recognized and handled during the course of such a program. 

For specific examples, let us turn to the Indianapolis Center for Advanced 
Research (ICFAR), where this concept of advanced research is practiced. One of 
the most interesting projects underway at the present time is early detection and 
treatment of breast cancer. Not only is breast cancer a major killer of women na- 
tionally, but it is a kind of death that is a very unhappy one; full of pain, cost, and 
emotional trauma. This problem should be directly approached, in my view. The 
research war on cancer that started during the Nixon administration; and, is 
presently funded through the Department of Health and Human Services and the 
National Cancer Institute, pays little heed to surgical techniques. Surgery is the 
present treatment for many forms of breast cancer. What we have suggested at 
the Center is an approach in which we employ ultrasound for early detection and 
then use focused ultrasound as a non-invasive surgical technique. We have made 
significant accomplishments in early detection of breast cancer using ultrasound 
through the support of the Showalter Residuary Trust. A small group of lay in- 
dividuals in Indianapolis were willing to listen to our request for funds to directly 
attack this major problem. The National Cancer Institute, with its concepts of 
basic research and generation of knowledge that would be put on the shelf, is not 
responsive to this kind of approach. The Showalter Trust decided to support a 
major program to attack this problem some years ago. We defined the results 
desired as (1) an increase in the probability of early detection, (2) a use of ultra- 
sound techniques to alleviate the normal radiation damage of mammography; and, 
(3) to provide a relatively low cost, rapid scanning device for wide-spread use. The 
barriers to the application of this kind of research, again, are numerous. First, 
there is the economic barrier to accomplish the instrumentation, the clinical 
testing, and all of the other technical work in association with such a project. For 
instance, let me suggest to you that after a major instrumentation company has 
laid out tens or hundreds of millions of dollars on the development of x-ray com- 
puterized tomography equipment, they do not relish the idea of a product coming 
on the market which will cost a tenth as much as their equipment and which is far 
more capable of early detection. Thus, it is difficult to find an avenue by which to 
commercialize the results of this type of work because of that kind of competitive 
situation. Another major barrier to the application of these techniques is the need 
for training of physicians. Of course, most radiologists and doctors have been in 
practice for a long time. You cannot depend upon medical schools to train physi- 
cians in ultrasound or you would be waiting for a decade to bring about the ap- 
plication of these techniques. It is not a simple matter to bring new instrumenta- 
tion and techniques to bear because of the need for extensive familiarization on 
the part of the physicians who will actually bring about the application of the ef- 
fort, not to mention technicians, nurses, and other medical personnel. The advanced 
research approach to this situation has been to develop the necessary instru- 
mentation in leased space in a hospital environment working closely with radio- 
logists and other medical technologists throughout the project. At the same time 
a small company, Medrix, Inc., was formed in order to market the resulting 



56 



Indiana Academy of Science 



system. The diagnostic phase of this effort is now nearing production having been 
demonstrated to be effective (see Figure 6). 




Figure 6. Prototype of Ultrasound Breast Scanning System 

Another example of advanced research is being conducted by the ICFAR 
Urban Systems Engineering Department. These scientists recognized, during an 
early study of energy usage in the City of Indianapolis, that a major problem 
existed in terms of energy consumption at the Belmont Street incinerator plant. 
As we looked further into this waste disposal plant, it was recognized that not 
only were there major costs involved in energy consumption; but, also, in replace- 
ment of critical parts. Therefore, a study was initiated in conjunction with the 
City to thoroughly instrument the incinerator plant to determine where losses 
were occurring and how improvements could be made. Prior to commencing the 
study, the results required were defined as (1) major fuel reduction, (2) stack emis- 
sion reductions so as to better meet EPA air quality requirements; and, (3) 
operating net reductions. A running incinerator plant was used as a test bed to 
obtain new information with respect to the operation of sludge incineration plants 
(see Figure 7). This research was successful and a solution was recommended in- 
volving a relatively simple feedback control system in which the gas composition 
of the exhaust stack controlled air flow to the incinerator. The resultant relatively 
constant temperature will not only reduce fuel flow, but also increase incinerator 
part life. This type of operation was successfully demonstrated for three days con- 
tinuously in March of 1978. This demonstration not only showed lower operating 
costs but also demonstrated emissions low enough to meet EPA standards. Per- 
manent changes are being made in the operation of the plant, which will bring 
about a reduction in cost of as much as $300,000 annually. Further, a higher opera- 
tional reliability can be attained, thus, reducing the requirements for future 
capital investments on the part of the City. In this program, scientists and 
engineers from ICFAR worked closely with supervision and workers at the in- 
cinerator plant itself, just as in the breast cancer detection project they worked 
with radiologists and hospital personnel. In this way, much of the not-invented- 



Presidential Address 



57 




Figure 7. Combustion Scientists Review Urban Sludge Incinerator Instrumen- 
tation 



here kind of attitude on the part of the practitioner is overcome because they are 
part of the development program. Finally, a small company was formed which will 
offer this service to other cities throughout the nation. 

As a final example of advanced research, let me present a logical approach to 
addressing the energy problem. First the major research results required in order 
to become more independent of foreign oil are: (1) conservation — methods to 
reduce energy usage, (2) exploration enhancement — techniques to reduce costs 
and time in finding new fossil fuels; and, (3) development of alternative fuels. In 
each of these cases, rapid strides can be made through application of the tech- 
niques of advanced research. ICFAR efforts take the following form: 

(1) Research on infiltration losses in fenestration in conjunction with 
manufacturers of storm windows and conducted in the housing environment with 
the aim of reducing cost of high thermal flow resistant storm windows for retrofit 
applications (see Figure 8). 

(2) Development of computer algorithms for tomographic reconstruction of 
underground regions by means of seismic, electromagnetic, or other signals 
through study of present fossil fuel systems in conjunction with producing com- 
panies. 

(3) Develop solar/gas hot water heating systems in conjunction with a gas 
utility for purposes of financing and maintenance arrangements. 

Each of these efforts has involved a licensing agreement with a small com- 
pany to bring about commercialization of the results. 

There are many other projects and programs at ICFAR, but all of them 
follow this general approach of advanced research. This is a type of research in- 
volving discovery, new techniques, and innovation, but the sense of discovery oc- 
curs in the course of the accomplishment of programs which relate to the future of 



58 



Indiana Academy of Science 




Figure 8. Thermal Resistance Measurement System 



the community and the nation, and are conducted in a style such that barriers to 
the applications of the research will be reduced as much as possible as the 
research results become available. The two cultures which C. P. Snow discussed 
so well are being brought together in an efficient manner at the working level. 



Presidential Address 59 

In summary, steps necessary for alleviation of economic problems according 
to this scenario are: 

1) Encourage venture technology capital. 

2) Apply principles of advanced research in developing public/private sec- 
tor research projects. 

3) Take time constants into account in assessing results. 

4) Support the scientists and the environment in which new ideas will con- 
tinue to flow. For with the scientist, lies the source of the intellectual property 
that will eventually benefit society. 

Literature Cited 

1. Armed Services Procurement Regulations, (ASPR), Section IV; 4-101, 
Definitions. 

2. Department of Labor, Bureau of Labor Statistics, Office of Productivity 
and Technology, "Comparative Real Gross Domestic Product, Real GDP per 
Capita, and Real GDP per Employed Civilian, Seven Countries, 1950-77," 
June 1978, Science Indicators, 1978. 

3. Department of Labor, Bureau of Labor Statistics, Office of Productivity 
and Technology, "Output per Hour, Hourly Compensation, and Unit Labor 
Costs in Manufacturing, Eleven Countries, 1950-77," November 29, 1978, 
Science Indicators, 1978. 

4. Flender, John 0. and Richard S. Morse, "The Role of New Technical Enter- 
prise in the U.S. Economy," MIT Development Foundation Study, 1979. 

5. Georgescu-Roegen, Nicholas. The Entropy Law and the Economic Process. 
Harvard University Press, Cambridge, Massachusetts, 1971. 

6. Gilder, George, "Should We Sacrifice Our Future to Preserve the Past?", 
INC., November 1980, pp. 93-98 (Portions of this article previously appeared 
in Harper's). 

7. Gilman, John J., "Inventions and Corporate Size," Physics Today, October 
1980, p. 9. 

8. IEEE Spectrum, February 1980 (Proposal: A National Engineering Founda- 
tion by Bruno 0. Weinschel) ICFAR extended this graph through 1979. 

9. Rifkin, Jeremy and Ted Howard. Entropy. Viking Press, New York, 1980. 

10. Science Indicators — 1978, National Science Board, National Science Foun- 
dation, 1979, p. 6. 

11. Snow, C. P. Two Cultures and the Scientific Revolution. Cambridge Univer- 
sity Press, 1959 (The Rede Lecture, 1959). 




Photo by Lloyd Anderson, IAS 



Robert E. Henderson 



Robert E. Henderson checks the projector and transparencies for use with his 
Presidential Address, "Scientific Research and Economic Indicators. " 



60 




Photo by Lloyd Anderson, IAS 

President Robert E. Henderson and Richard L. Conklin, Chairman of the Fellows 
Committee, greeted these distinguished members of our Academy at the annual 
banquet. These eleven members represent persons who have been Fellows of the 
Academy for at least twenty years. Numbers in parentheses indicate the year in 
which each person was elected a Fellow. 

Front Row, Left to Right: Richard L. Conklin C63), Willis H. Johnson C50). William 
A. Daily (W, Winona H. Welch ('35), Fay K. Daily f'58), and Robert H. Cooper ('55). 

Back Row, Left to Right: Alton A. Lindsey C50), Harry G. Day C-53), Ernest E. 
Campaigne C5U), Robert E. Henderson C79), Benjamin Moulton C53), Ira Baldwin 
C53), and William W. Bloom ('57). 

At the time of this photograph, this group collectively represented 508 years of 
membership of which 336 years (66%) were served as Fellows. 



61 




Photo by Lloyd Anderson, IAS 

Winona H. Welch 
One of the highlights at the annual banquet at St. Joseph's College was the 
recognition of members of the Academy who have been Fellows for at least twenty 
years. Winona Welch, who received special recognition from President Robert 
Henderson, set a record for the group by being a Fellow for forty- five years. 



62 



PROGRESS IN RESOLVING FOOD SAFETY PROBLEMS: SYSTEMATIC 
EVALUATION OF GRAS FOOD INGREDIENTS 



Harry G. Day,* Department of Chemistry, Indiana University 
Bloomington 47405 

A purpose of this discussion is to give some illumination to the status of food 
ingredients known as GRAS. The word is an acronym for the term Generally 
Recognized as Safe. Of necessity the discussion will include my own participation, 
with others, in a comprehensive and systematic evaluation of the safety of the 
several hundred GRAS food ingredients that may be used in food such as the 
many listed on cereal boxes, bread wrappers, cans of soup, and other food con- 
tainers. 

In connection with this large scale and timely process there is reason to make 
reference to the problems created by the prevalence of pseudoscience and 
zealotry in matters of food processing, food additives, and concepts of nutrition. 
That false concepts and confusion abound, and affect the resolution of problems 
concerning food processing and food additives, should be understandable because 
a disturbing proportion of the public is misinformed and lacks basic knowledge 
concerning science and nutrition (1). Thus there is wisdom in the saying of the 
once-noted humorous Josh Billings that "It's better to know nothing than to know 
what ain't so." 

Some of the current problems are not new. Long ago there was misinforma- 
tion and flagrant and extensive practices of food adulteration, misbranding, and 
use of certain food additives scarcely without regard for their effects on health. 
Apparently the first protective law of note on food was by King John (of Magna 
Carta fame) in 1202. This was the Assize (ordinance) of Bread (2). 

With the development of analytical chemistry in the last century it became 
possible to greatly enhance the detection of fraud in food processing. In the first 
quarter of that century the most noted consulting chemist and writer on food pro- 
cessing and adulteration was Fredrick Accum (1769-1838). 

The best known of Accum's books was the work on adulteration of food in 
1820 which became known as "Death in the Pot." The full title was "Treatise on 
the Adulterations of Food and Culinary Poisons, Exhibiting the Fraudulent 
Sophistications of Bread, Wine, Spirituous Liquors, Tea, Coffee, Cream, Pickles, 
and Other Articles Employed in Domestic Economy, and Methods of Detecting 
Them." As pointed out by the biographer of Accum, C. A. Brown (3), "This book is 
also a classic as it represents the first serious effort to cope with the difficult 
problem of food adulteration." It was a landmark in the beginning of the pure food 
movement. The action reached critical mas^ in this country in 1906 with the 
passage of the original Food and Drug Act. 

The general commitment to food safety in the government was headed for 
more than 25 years by Harvey W. Wiley (1844-1930), a native Hoosier who was the 
first professor of chemistry at Purdue. 

In 1902 Dr. Wiley inaugurated the first federal program "to investigate the 



'Indiana Academy of Science 1980-81 "Speaker of the Year" 

63 



64 Indiana Academy of Science 

character of food preservatives, coloring matters and other substances added to 
foods." By current standards his organization and program for inquiry, as a scien- 
tific undertaking, would not gain the approval of the most lenient of any granting 
agency for research. For example, Dr. Wiley employed 12 men who served as his 
"poison squad." His general procedure in testing the safety of certain food ad- 
ditives was to restrict all 12 to the same diet for a short time, with half receiving 
one compound and the others being given the same substance in a different 
form — or an entirely different compound. The appearance and feeling of the men 
were observed, and some limited metabolic studies were made. Notably, ex- 
perimental animals were not used. The testing program was largely employed for 
its publicity value. On that basis it had great effect. It focused attention on the 
need for actions to promote safety in food processing (4). 

Of course some chemicals used as preservatives or for other reasons were 
clearly harmful and measures to prohibit or properly control their use gradually 
became effective. But it was not until the middle of this century that explosive 
changes occurred which enormously expanded the application of myriad chemicals 
in the food industry. With the advent of new pesticides and other chemicals that 
could get into the food chain, and the elevation of environmental concerns such as 
were expressed by Rachel Carson in Silent Spring in 1962, significant numbers of 
aroused people vigorously sought ways to avoid food that contained residual or 
added chemicals. 

Although there was a basis for concern, pseudoscience and zealotry, as well 
as outright deception, soon mounted and there was a pyramiding of interest, much 
of it misguided, in the avoidance of chemicals in food. Many individuals and groups 
became fascinated by the idea of using so-called natural foods and organically 
grown foods. 

So-called health food stores became economically successful enterprises and 
the demand for "chemical-free" foods became felt in various segments of the 
economy. Since 1970 the growth of health food sales has risen an average of 30 
percent a year. It has caused major food manufacturers to consider acquiring 
health food businesses. Various large and well known food companies find it finan- 
cially advantageous to emphasize the naturalness of their products. 

The misdirected enthusiasm for so-called natural foods could be corrected 
through much better understanding of simple chemical facts about foods and the 
principles of nutrition. Commonplace foods such as oranges, beans, and potatoes 
contain naturally scores of identifiable chemicals some of which are the very ones 
that some natural food proponents abhor. Through advertising and in other ways 
some of the chemical manufacturers and food processors are giving some effort to 
the correction of public misconceptions. 

There is great justification for the widespread use of many food additives, in- 
cluding many classified as GRAS. Some reasons for their use are the following: 

1. To enhance and maintain desired consistency. Examples include 
lecithin, methyl cellulose, and mono- and diglycerides. 

2. To enhance nutritive value. Examples include various vitamins and 
inorganic salts. 

3. To enhance flavor. Examples include amyl acetate, benzaldehyde, 
ginger, sodium chloride, and monosodium glutamate. 

4. To control acidity or alkalinity. Examples are sodium bicarbonate, 
vinegar, citric acid, and lactic acid. 

5. To maintain appearance, palatability, and wholesomeness. Examples in- 



Speaker of the Year 65 

elude calcium propionate, ascorbic acid, and sodium benzoate. 

6. To give desired and characteristic color. Examples include carotene, 
chlorophyll, and many other natural and unnatural dyes. 

7. To mature and bleach. Examples include potassium bromate and iodate, 
hydrogen peroxide, and chlorine dioxide. 

8. Other functions. These include moisturizing in some foods and to pro- 
mote free-flowing in others. Examples respectively are glycerol and 
magnesium carbonate. 

In this setting it is timely to consider the large current program on the 
evaluation of food ingredients identified as GRAS. The listing of such substances 
began in 1958 in response to major amendments that year in the Food, Drug, and 
Cosmetic Act. The amendments constituted evolutionary changes in a process 
that started with the passage of the original Food and Drug Act in 1906 (5). 

Administration of the original Act was the responsibility of the Bureau of 
Chemistry of the Department of Agriculture. By 1958 the Food and Drug Ad- 
ministration (FDA) had been evolved from the Bureau and it was a part of the 
Department of Health, Education and Welfare, now Department of Health and 
Human Services. The GRAS ingredients are under the surveillance of the FDA. 
However, some jurisdictional complications exist in food additivies. The Depart- 
ment of Agriculture has a regulatory role concerning substances added to meat 
and other products of animal origin, and the Department of Interior has such 
responsibility concerning marine food products. There are even other federal 
regulatory agencies concerning the safety of food ingredients. 

Until 1958 the food safety laws were rather simplistic. If a substance was ap- 
parently safe and nondeceptive in its use it was permitted, but if it was regarded 
to be "poisonous and deleterious" it was prohibited. The important differences 
made by the quantity ingested were not given sufficient consideration in the law. 
By 1958 the young science of toxicology and the status of analytical chemistry had 
advanced far enough to make rational and appropriate administration of the law 
impossible. It was necessary to understand that the possible beneficial effects or 
harmful effects had to be a function of the amount of a substance ingested. Even 
Paracelsus had recognized this in the 16th century, when he wrote, "it is only the 
dose which makes a thing a poison." That principle had to be recognized in 
establishing and implementing the new food safety laws. 

The Food Additives Amendment required the fixing of the responsibility for 
demonstrating safety of the food ingredient on the industrial firm proposing its 
use in interstate commerce. However, for practical reasons the Act specifically ex- 
empted food additives in common use at that time which were generally recogniz- 
ed as safe as determined by "experts qualified by scientific training and ex- 
perience," or "experience based on common use in food." 

The first listing of generally recognized as safe additives was prepared by 
FDA scientists and without benefit of substantial input from other scientists. 
Shortly thereafter approximately 900 scientists in the academic world, industry, 
and government were requested by direct mailing to evaluate the list and suggest 
additions and deletions. The mailing list was prepared by reference to American 
Men of Science (at the time women were not recognized in the title) and several 
professional societies. I recall that I was invited to participate. 

During the 1960s there were several changes in the GRAS list including 
those resulting from direct authorizations by letter from the FDA to food 
manufacturers or processors. Such statements of opinion or authorization became 



66 Indiana Academy of Science 

known as "GRAS letters." I am not aware of any inappropriate decisions through 
this process of dealing with the Food Additives Amendment of 1958. 

Action leading to a thorough evaluation of GRAS substances was called for 
by President Nixon in 1969. Almost concurrently the White House Conference on 
Food, Nutrition, and Health was convened (6). The Conference concluded that 

"Traditional or long-continued use of any additive can no longer be 
considered to be sufficient evidence of safety. Thus it is necessary that a 
continuing re-evaluation be maintained of all compounds whose use in 
foods is relatively freely allowed." 

The Conference "Recommended that the list of substances known as GRAS be 
systematically reviewed for safety in the light of new knowledge, experience, new 
levels, and new categories of food use." 

Within the same general time period the need for changes in GRAS was 
manifested through mounting consumer activism, the media-stimulated fright 
over the use of cyclamates as sugar substitutes, and other media-produced alarms 
on the status of several GRAS substances. 

The eventual action of consequence by the FDA was the establishment of a 
contractual arrangement with the prestigious and long established Federation of 
American Societies for Experimental Biology (FASEB) through which a com- 
prehensive assessment of a large proportion of GRAS food ingredients would be 
conducted (7). The Federation, through its Life Sciences Research Office, 
established a committee and staff to carry out the large project. I became a 
member of the committee, the Select Committee on GRAS Substances (SCOGS), 
in 1973. The Committee was assisted through other contracts made by the FDA to 
provide extensive searching and compilation of the world's literature on each of 
several hundred GRAS food ingredients. Other assistance came from: (a) a con- 
tract with the National Academy of Sciences-National Research Council for 
surveys to estimate the human consumption of GRAS food ingredients, and (b) 
from other organizations to provide special information including mutagenic and 
teratogenic testing of relevance to the Select Committee. 

The original Select Committee was composed of nine research scientists 
representing diverse backgrounds of importance. From 1972 through 1980 there 
were two resignations and four additions to the Committee. The expertise during 
most of the time included recognized persons in foods and nutrition, biochemistry, 
pharmacology and toxicology, pathology, and medicine including pediatrics and 
oncology. 

An important consideration in the selection and functioning of the Committee 
was to assure professional competence and objectivity. To avoid even the ap- 
pearance of any conflict of interest it was necessary for Committee members to be 
free from significant affiliation with industrial firms involved with food, govern- 
mental agencies, or "consumer-oriented" organizations with probable bias 
concerning GRAS food ingredients and the enforcement of food safety regula- 
tions. Initially there was considerable agitation from a limited sector of the public 
for the inclusion of consumer-oriented representation on the Committee. Capitula- 
tion to the effort obviously would have compromised the principle of high objec- 
tivity and limitation of activity to scientific evaluation of GRAS substances. The 
agitation subsided after two or three years. 

A vital component of the system for evaluation was the supporting staff. 
During most of the program approximately six full time professional staff scien- 



Speaker of the Year 67 

tists gave a substantial proportion of their time to the work of the Select Commit- 
tee and they were assisted by several competent and responsible secretarial 
workers. Even though the staff personnel were not responsible in the formulation 
of Committee opinions and conclusions, all were required to be without conflict of 
interest. Even persons who conducted the extensive literature surveys were pro- 
hibited from expressing opinions on GRAS ingredients in the compilations of the 
findings. 

To maximize independent and searching evaluation of basic information each 
initial draft report was written by Committee members assigned so as to match 
subject matter with the professional backgrounds of the individuals. Such drafts 
were individually reviewed by all members and then discussed in full committee 
meetings attended by the professional staff members. A serious attempt was 
made to gain input from all areas where searching inquiry and evaluation might 
be useful. In dealing with several substances scientists with special knowledge 
and background were consulted. Committee discussion and inquiry on a topic was 
continued at the initial meeting, or at subsequent meetings, until there was agree- 
ment that the draft content, opinion, conclusions, and available supporting data 
reflected adequately the views of each member. 

At this point a new draft report was prepared by the professional staff and 
after thorough processes of individual review and criticism in the Committee a 
final draft report was written in which all statements and data were verified 
against the original articles and other sources. Such drafts upon signed approval 
by the Committee members became the tentative report of the Select Committee. 

The tentative reports, after approval by an advisory committee of the 
Federation, were transmitted to the FDA. Announcements of the availability of 
the reports to the public were always made in the Federal Register along with in- 
vitations to request a public hearing or furnish the Committee with data, informa- 
tion or views (8). When requested, public hearings were held and submitted writ- 
ten data, information, or views were accepted and considered. The Select Commit- 
tee submitted 143 final reports to the FDA. Table 1 lists representative GRAS 
food ingredients evaluated by the Committee. 

Table 1. Representative GRAS Food Ingredients Evaluated by the Select 
Committee on GRAS Substances* 

Gum arabic Ascorbic acid and various ascorbates 

Butylated hydroxytoluene (BHT) Magnesium salts 

Benzoic acid and sodium benzoate Silicates 

Sorbitol Sucrose 

Sulfiting agents Bioflavonoids 

Nutmeg, mace, and their essential oils Propionates 

Certain zinc salts Pectin and pectinates 

Alginates Caffeine 

Phosphates Vitamin D, vitamin D 2 , and vitamin D, 

Iron and iron salts Pyridoxine and pyridoxine hydrochloride 

Certain glutamates Potassium chloride and sodium chloride 

Protein hydrolyzates Lecithins 

Choline chloride and choline bitartrate Niacin and niacinamide 

Aluminum compounds Thiamin, thiamin hydrochloride, and thiamin mononitrate 

Certain calcium salts Carotenes 

Dextrose, corn syrup, and invert sugar Riboflavins 

Butylated hydroxyanisole (BHA) Activated carbon (charcoal) 

Gelatin Vitamin A, vitamin A acetate, and vitamin A palmitate 

*Copies of all the final reports may be purchased from the National Technical In- 
formation Service, Springfield, VA 22161. Order number, PB 80203789 



68 Indiana Academy of Science 

The heart of the reports were the opinions and the conclusions. These were 
derived from the comprehensive analysis of the literature made by the Committee 
and reviewed in the body of the reports. 

It was possible to reduce the basic conclusions to five categories as follows: 

1. The food ingredient should continue in GRAS status with no limitations 
other than good manufacturing and handling practices and levels of use 
that might reasonably be expected in the future. 

2. Same as No. 1 except that without additional supporting data the level 
of use should not be increased. 

3. Same as No. 1 except that uncertainties exist requiring that additional 
studies be promptly conducted, and evaluated. 

4. There is evidence of adverse effects and safe usage conditions should be 
established or the GRAS status should be rescinded. 

5. There is insufficient information upon which to base an evaluation. 

A large majority of the GRAS substances were found to present no hazard to 
health when used at levels that are now current and in the manner practiced. 
Substances in this category include sodium benzoate, calcium salts and certain 
other inorganic salts, and all the vitamins except A and D. 

Substances recommended to be restricted to the current level of use include 
vitamins A and D, glutamates, zinc salts, propionates, hydrogen peroxide, and 
many gums such as gum arabic, and gum ghatti. 

Several GRAS substances were found to be questionable when used at levels 
that are now current and in the manner practiced. These include oil of nutmeg, 
BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene), and caffeine. 

Sodium chloride is the only GRAS ingredient of widespread interest which 
the Committee concluded is being used at such high levels, both discretionally and 
in processed foods, that the health of a significant proportion of the people is be- 
ing jeopardized. 

Several food ingredients were considered concerning which there is insuffi- 
cient information upon which an evaluation can be made. In general there is little 
if any commercial interest in using any of these substances in food. 

Benzoic acid and sodium benzoate were among the earliest GRAS ingre- 
dients reviewed by the Select Committee. These long-used food preservatives 
were found to be without evidence of being harmful when used at current levels 
or that might reasonably be expected in the future. It is of interest that Harvey 
Wiley had fed these compounds to his "poison squad" at the turn of this century. 
Although his testing gave no evidence of harmfulness, he persisted in his belief 
that they should not be used in food. 

The evaluation of 415 GRAS ingredients was time-consuming and difficult for 
the Select Committee. Some that required the greatest amounts of time included 
caffeine, sucrose, glutamates, sodium chloride, and iron and iron salts. What were 
some of the considerations and actions? This may be illustrated by a discussion of 
caffeine (9). 

It is common knowledge that caffeine is a prominent constituent of coffee 
and tea. Also, it is present, as required by the Code of Federal Regulations, in all 
cola-type beverages. However, the caffeine concentration in such beverages must 
not exeed 0.02 percent by weight. Less than 10 percent of the caffeine present in 
these beverages is from extracts of kola nut; all the remainder is added caffeine. 



Speaker of the Year 69 

It is not clear whether caffeine is added to cola-type beverages for its stimulatory 
effects or for the enhancement of flavor. The substance is without nutritive value. 

In its evaluation of caffeine the Select Committee focused attention on its use 
as commercially added to food and beverages. Coffee, tea, and cocoa are not in the 
GRAS category, but their contribution to the total consumption of caffeine was 
considered. 

It is established that the dose of caffeine required to stimulate central ner- 
vous system activity in humans is approximately 3 mg per kg of body weight and 
the effect is observable at about 2 mg per kg. A substantial proportion of cola 
drinkers consume about 0.3 mg of caffeine per kg per day from this source with a 
few in«the less than 5 year age range consuming as much as 1.8 mg per kg per day. 
As stated in the Select Committee's report on caffeine, "the consumption of a 
12-ounce container of cola beverage containing 0.01 percent caffeine represents a 
dose of about 0.9 mg per kg for a 40 kg child, for example, or about 0.6 mg per kg 
for an adult. It is to be noted that these figures represent the amount of caffeine 
solely from cola drinks." 

Another consideration is the possible adverse effect of frequent selection of 
cola drinks and other soft drinks in preference to milk, particularly when the diet 
is marginal in the nutrients which milk abundantly supplies. 

The Select Committee concluded that "it is not appropriate to continue to 
consider caffeine as a generally recognized as safe substance for addition to cola- 
type beverages." It pointed out that in support of this conclusion "The amount of 
caffeine consumed as cola-type beverages borders on the dose known to produce 
central nervous system stimulation in animals and man. Whether such stimulation 
constitutes an adverse effect or whether a potential hazard may exist for the seg- 
ment of the population, particularly children, that is exposed to stimulating doses 
of caffeine, cannot be answered on the basis of the evidence available." 

In response the FDA proposed regulations that would encourage the produc- 
tion of caffeine-free cola beverages and require the industry to conduct new caf- 
feine studies. 

Prior to the response the Bureau of Foods of the FDA conducted a study 
which indicates a significant proportion of birth defect in the offspring of female 
rats that had ingested relatively large quantities of caffeine. 

A representative press release, by the Associated Press, in October 1980 
stated the following (10): 

"One of the regulations put forward by the Food and Drug Ad- 
ministration would make the continued use of caffeine as a food additive 
contingent upon the industry's funding of studies showing how caffeine 
affects children and human fetuses. 

"Both the National Coffee Association and the soft drink industry 
already have indicated willingness to pay for additional research. 
Among the studies they propose is one comparing rates of birth defects 
with caffeine consumption by pregnant women. 

"The other proposal would remove caffeine from the agency's list 
of substances generally recognized as safe, a step that ordinarily would 
lead to prohibition of its use (as an additive). The agency would allow an 
exemption permitting the current uses of caffeine to continue while the 
studies are conducted." 



70 Indiana Academy of Science 

Throughout the long period of evaluation by the Select Committee and the 
subsequent deliberations and limited research by the FDA, the Center for Science 
in the Public Interest acted vigorously to promote the limitation of caffeine con- 
sumption among children and pregnant women. This consumer-oriented action 
group has strongly criticized the FDA for not going further in withdrawing caf- 
feine from the GRAS list. The Center wanted the limitation to start at once, on 
the basis of the present evidence, and it wanted the labeling of coffee to be in- 
itiated with a warning that would advise pregnant women to avoid it during 
pregnancy. I believe the FDA's response to the Select Committee's report was 
correct. 

Moderate use of coffee, tea, and cola drinks by adults not under stress 
presents no cause for concern, if such use does not result in the exclusion of milk 
or other food that may be needed. Granted that caffeine has a clearly stimulating 
effect on the central nervous system at about 3 mg per kg of body weight, an 
adult weighing 70 kg, or about 150 pounds, should be cautious about ingesting 
more than approximately 200 mg per day. At 85 mg of caffeine per cup of coffee 
the consumption of more than about 2 cups of coffee, a cup of tea, and a cola drink 
or two per day can be assumed to be disadvantageous. 

In retrospect, over the years that the Select Committee reviewed, evaluated, 
discussed, and worked toward the completion of the reports on caffeine and many 
other GRAS food ingredients, I believe there was no improper action or in- 
terference of any kind by the advocates of change and the advocates of the status 
quo. 

From the perspective of 1980-81 the methodology and guidelines used to 
evaluate the GRAS ingredients have stimulated new concepts and actions in the 
assessment of food safety (7). 

It is hoped that this will be followed in other problem areas where the objec- 
tivity and methodology of science can be used in providing information and judg- 
ment through which public decisions can be made. This could be applicable at the 
state level as well as the national level. I believe the talent in the Indiana 
Academy of Science should be utilized in studying and reporting on some of the 
problems important in this State. 



Literature Cited 

1. Barrett, S. and G. Knight. 1976. The Health Robbers. George F. Stickley 
Co., Philadelphia. 

2. Bernarde, M. A. 1971. The Chemicals We Eat. McGraw-Hill, New York, p. 
163. 

3. Brown, C. A. 1925. The life and chemical services of Fredrick Accum. Jour. 
Chem. Educ. Vol 2, 1008-1034. 

4. Anderson, 0. E., Jr., 1958. The Health of a Nation. University of Chicago 
Press (Harvey W. Wiley and the Fight for Pure Food) 1975. 

5. Hall, R. L. 1975. GRAS -Concept and application. Food Technol. Vol. 29, p. 
48-53. 

6. White House Conference on Food, Nutrition and Health. Final Report, 
December 24, 1969. U. S. Government Printing Office, Washington, D.C. 

7. Siu, R. G. H., J. F. Borzelleca, C. J. Carr, H. G. Day, S. J. Fomon, G. W. 
Irving, Jr., B. N. LaDu, Jr., J. R. McCoy, S. A. Miller, G. L. Plaa, M. B. 
Shimkin, and J. L. Wood. 1977. Evaluation of health aspects of GRAS food in- 



Speaker op the Year 71 

gredients: lessons learned and questions unanswered. Federation Pro- 
ceedings, Vol. 36, p. 2519-2562. 

8. Department of Health, Education, and Welfare. Food and Drug. 1974. 
Food additives and GRAS Substances. Federal Register. Vol. 39, number 
185, Part II, p. 34172-34219. 

9. Select Committee on GRAS Substances. 1978. Evaluation of the Health 
Aspects of Caffeine as a Food Ingredient. SCOGS-89. Report to Food and 
Drug Administration from Life Sciences Research Office, Federation of 
American Societies for Experimental Biology, 9650 Rockville Pike, Bethesda, 
Maryland 20014. 

10. News Item by Associated Press. 1980. Food, drug agency is working toward 
caffeine-free nation. Indianapolis Star, 22 October 1980. 



ANTHROPOLOGY 

Chairman: Francis X. Grollig 

Department of Anthropology, Loyola University, Chicago, 

Illinois 60626 

Chairman-Elect: Charles P. Warren 

Department of Anthropology, University of Illinois 

at Chicago Circle, Chicago, Illinois 60680 



ABSTRACTS 

Details in Mexican Archaeology. Francis X. Grollig, S.J., Department of An- 
thropology, Loyola University of Chicago, Chicago, Illinois 60626. With the 

aid of a small grant from Loyola University it was possible to do extensive work 
in photographing details of larger artifacts and architectural remains in a series of 
some 18 archeological zones visited in the (third) annual Loyola Program in Mex- 
ican Art and Archeology this (1980) summer. In this presentation we will see 
selections from Calixtlahuaca, Cuicuilco, Dainzu, Mitla, Monte Alban, Tenayuca, 
Teotenango, Teotihuacan, Tlatelolco, Tula, Xochicalco, Yagul and the Coyolxau- 
qui. 

The Indiana Packet: Ecotone and Cultural Boundary within the Lower Wabash 

Valley. C. Dean Higginbotham, Owensville, Indiana 47665. Deam (1940) in his 

synthesis of Indiana flora identified a unique ecological setting in the 
southwestern tip of Indiana which he named the "Indiana Pocket". The Indiana 
Pocket is characterized by typically southern species of plants and animals, which 
are at their northern limits in the lower Wabash Valley. Archeological evidence 
indicates that the Indiana Pocket was not only a boundary for plant and animal 
communities but also for human communities as well. 

Cataract Chert And Its Distribution In West-Central Indiana. Robert E. Pace, 
William Rheinhardt, and Daniel Thiel, Indiana State University, Terre Haute, 

Indiana 47809. A description and discussion of bedrock sources of Cataract 

Chert along Mill Creek and tributaries in Owen County and a discussion of the 
regional distribution of the chert at archaeological sites in West-Central Indiana. 

Stone Box Burials In West-Central Indiana. Robert E. Pace, Indiana State 

University, Terre Haute, Indiana 47809. Three stone burial boxes, from sites 

in Parke, Vermillion and Sullivan counties are discussed, with comments on their 
cultural affiliation and their relationship to culture complexes to the south. 

A Note on the Distribution of Riverton Points. Curtis H. Tomak, Martinsville, 

Indiana, 46151. Howard Winters' definitive monograph on the Riverton 

Culture was published in 1969. Based upon data available to him at the time 
Winters presented a geographical distribution of Riverton materials, discussed a 
settlement pattern and settlement system, and suggested an area of origin for the 
Riverton Culture. Based upon additional data Riverton or Riverton-like materials 
are known to have a significantly wider distribution and to be common in areas 
other than that originally indicated for the Riverton Culture. This paper presents 
some data regarding this wider distribution; t\ 's has implications for interpreta- 
tions of Riverton. 

72 



Anthropology 73 

Rock Shelters: An Important Archaeological Resource of Southern Indiana. 

Curtis H. Tomak, Martinsville, Indiana, 46151. Rock shelters are numerous in 

some parts of southern Indiana. Since a large number of them were utilized by 
prehistoric peoples and since rock shelters can provide data relating to a variety 
of archaeological problems, they constitute one of the State's important ar- 
chaeological resources. Unfortunately, many rock shelters have been disturbed by 
artifact collectors thereby seriously impairing this cultural resource. It would be 
worthwhile to initiate surveys structured to locate and to assess the research 
potential of rock shelters before destruction of the data base proceeds much fur- 
ther and to do this within the context of a preservation program. This paper in- 
cludes a discussion of a survey which located several rock shelters. Two of them 
were excavated, found to be significant sites, and preserved by the State of In- 
diana. 

Two Case Studies in Forensic Anthropology: Calumet Township, Indiana; and 
Griffith, Indiana. Charles P. Warren, Department of Anthropology, University 

of Illinois at Chicago Circle, Chicago, Illinois 60626. In July and August 1979, 

two cases involving the recognition and identification of human remains were 
brought to the physical anthropology laboratory of the Department of An- 
thropology, University of Illinois at Chicago Circle. Both cases were from sites in 
northern Indiana, one from Calumet Township, Indiana, and the other from Grif- 
fith, Indiana. Discussion of the forensic problems inherent in each case and the 
means of resolving these problems reveals that the forensic anthropologist must 
use a comprehensive and eclectic approach and must draw upon related 
disciplines to provide logical answers to the questions which arise concerning 
unknown remains which have undefined temporal and spatial frames of reference. 



An Experiment with Time Control in Intensive Site Survey 

James 0. Bellis 

Department of Sociology and Anthropology 

University of Notre Dame, Notre Dame, Indiana 46556 

Acknowledgements 

The research for this study was conducted under the auspices of the Founda- 
tion for Illinois Archaeology and Northwestern University as a part of the pro- 
gram for the Advanced Field Methods course in the summer of 1974. I would like 
particularly to thank David Asch, Tom Cook and Ken Farnsworth whose sugges- 
tions were invaluable to the present project; the responsibility for its conduct, 
however, is my own. 

The last three decades have seen a rapid and systematic development of 
more rigorous controls in all aspects of archaeological method. The earlier efforts 
were directed toward improving excavation and data recovery and preservation 
techniques. These developments were followed by increasingly precise and com- 
plex methods of analysis of excavated material. Most recently, perhaps in part 
stimulated by the needs of cultural resource management, attention has been 
directed to the improvement of the methods of archaeological site reconnaissance 
and survey. 

The questions asked of site survey research are phrased at many different 
levels. Some questions have a broad regional focus with such goals as the deriva- 
tion of prehistoric settlement patterns. Others are phrased at the level of the 
assessment of the individual site, attempting to discover intrasite patterns such 
as specialized activity areas within a site. The present paper addresses itself to 
certain aspects of the intensive single site survey. 

Often the pre-excavation assessment of the surface debris scattered on a site 
is limited to the simple pedestrian "walk over" survey. In its least complex form 
this approach has the archaeologist spending enough time on the site to gain a 
sense of familiarity. If excavation is planned to follow the survey, then this in- 
tuitive familiarity will be the basis for placement of the excavation units. The 
result of such an informal method can range from excellent (in which case the 
archaeologist will be said to have a "good nose") to the disastrous (in which case 
the archaeologist will be said to have defined what must be done during the next 
field season). 

Many problems result from such an unstructured approach to site survey. 
The most obvious are the short range problems which come from having to base 
the excavation strategy of any site on such a haphazard body of data. Also 
resulting from such unsystematic site surveys are problems with long range im- 
plications. For example, the result of the original survey cannot be replicated 
even by the original investigator. Therefore it is impossible for other in- 
vestigators to either systematically follow the original research strategy at the 
same site or to conduct meaningful comparative studies at other sites. In short, 
the results of unsystematic surveys are incomparable. 

Two general strategies have developed as solution to the problems created 
by the unsystematic survey. The most obvious solution is literally to pick up 
everything present on the surface of a site. The primary methodological concern 
in applying this total "vacuum cleaner" approach is to systematize the pick up by 

74 



Anthropology 75 

some means of spatial controls (e.g. a grid system) so that the patterns of surface 
scatter can be reconstructed accurately in the laboratory. The other strategy for 
resolving the problems of the unsystematic survey is to select, by means of some 
systematic sampling procedure, a representative portion of the total site surface 
for survey. In this second approach, the area sampled also is treated 
characteristically with a total surface pick up. 

In either of these two approaches, enormous amounts of man hours are re- 
quired for the actual pick up operation. The present study demonstrates that a 
refinement of the actual pick up procedure by means of imposing a time control 
can increase greatly the efficiency of the use of the field labor crew without 
decreasing the accuracy of the sample or the replicability of the results. It is pro- 
posed that the first 30 minutes of surface pick up in small finite areas can be done 
in such a way that it will repeatedly produce data sets which are accurate 
representations of the data sets produced by total surface pick ups of the same 
areas. That is to say that once the sampling strategy has been chosen for a given 
site surface, it should be possible to gather in 30 minutes, a sample of surface 
material which is accurately representative from each of the surface units 
designated for pick up, and that therefore a total "vacuum cleaner" pick up is un- 
necessary. 

The Mound House Site, a Middle Woodland village site on the lower Illinois 
River valley, was chosen for a test site. This site is quite large, with over 40,000 
M 2 exposed by cultivation at the time of the survey. The amount and type of 
cultural material scattered on the surface of the site varied from one area to 
another. On the basis of a cursory walk over the surface of the site, five sub-areas 
were defined. Each of these sub-areas exhibited a distinct type and density of sur- 
face material present. Three of these were isolated zones of extremely dark 
organically stained soil which had very intense amounts of debris including large 
amounts of pottery, bone, fire-cracked rock and burned limestone. For purposes of 
this report these are labeled Areas 1, 2 and 3. Area 4 was a large Middle 
Woodland mound and the adjacent slope or apron of the mound down to the level 
of the original surface. This mound and its immediate environs exhibited the 
lowest amount of surface debris of any area within the site. Finally, the remaining 
portion of the site around and amongst Areas 1, 2, 3 and 4 uniformly exhibited a 
moderate scatter of chert and pottery, but showed very little organic staining, 
fire-cracked rock or burned limestone. This general area has been labeled Area 5. 
A test unit was placed in each of these five areas. 

To each one of these units, a survey crew member was assigned. Each crew 
member was instructed to collect a sample of the surface material which would, in 
his judgment, be an accurate representation of the total population of cultural 
material present in that unit. In conducting site surveys with both professional 
and student archaeologists, it has been my experience that the representative 
quality of the sample collected by most workers is increased as the size of the sur- 
face area to be covered is decreased. Therefore relatively small units (6M x 6M) 
were used in the present study. The size of the unit used is ultimately arbitrary 
but influenced by such factors as the nature of the terrain, the type of site, the 
condition of the site surface, and the general level of experience of the crew. 

Both instruction and supervision were important to the success of the 
method, and should not be slighted in the preparation of a crew. The idea of a 
representative sample was clearly explained, and an example chosen from another 
collection was illustrated in the laboratory. In the field the crew was instructed to 



76 



Indiana Academy of Science 



keep moving within their assigned 6M x 6M unit during the pick up periods, and 
cautioned not to "settle down" to exhaustively pick up any one part of the unit. 
These goals and instructions were repeated as often as necessary during the 
supervision of the actual field work. The actual pick up procedure was conducted 
in 30 minute intervals. At the end of each 30 minute pick-up period the bags of col- 
lected material were closed and labeled for the test unit and period. This process 
was repeated until the surface material from each of the 6M x 6M test units had 
been totally collected. 

The material was then separated in the laboratory into the following six 
gross classes of material: limestone, chert, pottery, fire-cracked rock, bone/shell 
and miscellaneous. For each test unit the collected material was cumulatively 
quantified by weight at the end of every 30 minute pick up period. These figures 
were then converted to a percentage value for each of the gross classes of 
material as a part of the total material collected. For example, by consulting Table 
1, Unit 1 the figures entered for the 90 minute period represent the cumulative 
total of each class of material collected in the first 90 minutes of pick up, stated 
both in terms of absolute weight and in terms of the percentage that each 
material class made up of the total collection. 

The results of this experiment are presented below in both tabular (Tables 
1-5) and graphic (Figures 1-5) form. An examination of this data demonstrates that 
in three (Units 1, 2 and 3) of the five units tested the relative proportions of the 
gross classes of material remained extremely regular from the first 30 minutes 
through the completion of the total pick up. It should be noted that these three 
units represent the areas with the most intense amount of materials present on 
the surface. It should also be noted that the units which produced the most erratic 
results, Units 4 and 5 were the areas of the lowest density of debris on the sur- 
face. It is suggested that this positive correlation between debris density and pat- 
terns of regularity in the collected surface material is due to the fact that in a low 
density area the difference of only a few pieces of material has an exaggerated 
statistical impact on the small body of data. It is further suggested that the prob- 
lem of these irregularities could be minimized by experimentally enlarging the 
pick up unit when the density of surface material is relatively low. 

The data are divided into gross classes of material and are presented in the 
following tables as both the absolute weight in grams then as a percentage of the 
total weight of all material collected at the end of each time period. Percentages 
are calculated to the nearest 0.5%. 

Table 1. (Area 1) 



time 














fire cracked 












elapsed 


limestone 


chert 


pottery 


rock 


bone/shell 




miscellaneous 




g- 


% 


g- 


% 


g- 


% 


g- 


% 


g- 


% 




g- 


°/o 


30 


1049 


19 


2722 


49 


687 


12.5 


737 


13.5 


298 


5.5 




43 


1 


60 


1517 


20 


3430 


45 


1042 


13.5 


1148 


15 


369 


5 




106 


1.5 


90 


1644 


19.5 


3771 


44.5 


1226 


14.5 


1304 


15.5 


411 


5 




128 


1.5 


120 


1729 


19 


4012 


44 


1453 


16 


1304 


14.5 


475 


5 




149 


1.5 


150 


1814 


19.5 


4111 


44 


1488 


16 


1304 


14 


496 


5 




168 


2 












FIGURE 1 (AREA 1) 














TIME 
















% FIRE-CRACKED 










ELAPSED 


% LIMESTONE 


% CHERT 




% POTTERY 


ROCK 




% BONE/SHELL % 


MISC. 


30 




i i 


49 L 






12.5 




13.5 i 1 












60 


20 
19. 


s" 


45 c 
44.5c 




— , 


13.5 
14.5 


[=> 


15.5 c=) 
14.5 c=] 
14 C=3 




5 
5 
5 

5 


□ 
□ 


1 


5 


90 






5 D 




— ' 




19. 


5| 1 


44 c 






16 


i 1 




□ 


2 




150 

























Ant 


HROPOI 


X)GY 










77 












Tabi 


.K2.1A 


rea2) 












time 














fire cracked 










elapsed 


limestone 


ch< 


>rt 


pot 


tery 


rock 


bom 


/shell 


misc< 


•llaneous 




g- 


°/o 


R- 


°/o 


g. 


% 


g- 


% 


R- 


% 


K- 


% 


30 


3274 


52.5 


1418 


23 


524 


8.5 


794 


13 


189 


3 


14 


.5 


60 


5316 


51.5 


2098 


20.5 


1006 


9.5 


1588 


15.5 


317 


3 


24 


.5 


90 


5493 


50.5 


2230 


20.5 


1097 


10 


1666 


15.5 


359 


3.5 


43 


,5 


120 


5939 


50.5 


2393 


20.5 


1225 


10.5 


1694 


14.5 


416 


3.5 


48 


.5 


150 


6322 


51 


2512 


20 


1374 


11 


1694 


13.5 


458 


3.5 


54 


.5 


180 


6443 


50.5 


2569 


20 


1452 


1 1 .5 


1751 


13.5 


501 


4 


60 


.5 


210 


6684 


50.5 


2588 


19.5 


1551 


12 


1751 


13.5 


548 


4 


65 


.5 


240 


6740 


49.5 


2723 


20 


1735 


12.5 


1821 


13.5 


548 


4.5 


71 


.5 



FIGURE 2 (AREA 2) 



TIME 
ELAPSED 

30 


LIMESTONE 


52. 5 [ i 




51 .5 | | 


120 






150 






180 






210 






240 







23 

20.5 

20.5 

20.5 

20 

20 

19.5 

20 





% FIRE-CRACKED 






1 POTTERY 


ROCK 


BONE/SHELL 


% 


8.5CH 


13 c=n 


3 D 


.5 


9.5CD 


15.5 c=) 


3 Q 


.5 


10 en 


15.5 n 


3.5 □ 


.5 


10.5 c=i 


14.5 1=1 


3.5 a 


.5 


11 □ 


13.5 □ 


3.5 a 


.5 


11.5i — i 


13.5 □ 


4 D 


.5 


12 en 


13.5 t=) 


4 D 


.5 


12.5 CZD 


13.5 C=n 


4.5 a 


.5 



Table 3. Urea 3) 



time 














fire- 


cracked 










elapsed 


limestone 


chert 


pottery 


rock 


bone/shell 


miscellaneous 




g- 


% 


g- 


% 


g- 


% 


g- 


% 


g- 


°/o 


g- 


% 


30 


992 


29.5 


1190 


35.5 


383 


11.5 


680 


20 


57 


1.5 


57 


1.5 


60 


1758 


35 


1758 


35 


517 


10 


872 


17.5 


85 


1.5 


62 


1 


90 


1871 


34.5 


1970 


36 


553 


10 


879 


16 


106 


2 


81 


1.5 


120 


2070 


35.5 


2065 


35.5 


602 


10.5 


879 


15 


120 


2 


87 


1.5 


150 


2145 


34 


2334 


37 


659 


10.5 


879 


14 


142 


2.5 


125 


2 


180 


2202 


34 


2398 


37 


702 


11 


888 


13.5 


163 


2.5 


139 


2 


210 


2301 


33.5 


2582 


37.5 


773 


11.5 


888 


13 


163 


2.5 


139 


2 


240 


2320 


33 


2674 


38 


810 


11.5 


888 


12.5 


170 


2.5 


148 


2 


270 


2356 


32 


2887 


39 


909 


12.5 


894 


12 


204 


3 


158 


2 


300 


2448 


32 


2958 


38.5 


938 


12.5 


900 


12 


238 


3 


172 


2 


330 


2476 


31.5 


3000 


38.5 


987 


12.5 


905 


11.5 


266 


3.5 


186 


2.5 


360 


2767 


32 


3255 


37.5 


1186 


13.5 


953 


11 


316 


3.5 


222 


2.5 












FIGUR 


3 (AREA 3) 













TIME 
ELAPSED 


% Lie 

29.5 

36 

34.5 

35.5 

34 

34 

33.5 

33 

32 

32 

31.5 

32 


EST0NE 


60 


1 ' 




90 






120 






150 






180 






210 






240 
270 




' ' 




300 






330 






360 







35.5 

35 

36 

35.5 

37 

37 

37.5 

38 

39 

38.5 

38.5 



37.5 c 



FIRE-CRACKED 






ROCK 


BONE/SHELL 


% MIS 


20 i 1 


1.5q 


1.5 Q 


17.5 , , 


1.5 B 


1 


16 ccn 


2 □ 


1.5 o 


15 cnn 


2 o 


1.5 o 


14 (=1 


2.5 o 


2 □ 


13.5 en 


2.5 a 


2 o 


13 cm 


2.5 


2 o 


12.5 en 


2.5 Q 


2 o 


12 C=D 


2.5 O 


2 a 


12 en 


3 o 


2 o 


11.5 en 


3.5 D 


2.5 


11 C=l 


3.5 D 


2.5 O 



78 




Indiana Academy 


of Science 














Table 4. (Area 4) 










time 








fire-cracked 








elapsed 


limestone 


chert 


pottery 


rock 




bone/shell 


miscellaneous 




g. % 


g. % 


g. % 


g- 


% 


«?. % 


g- 


% 


30 


64 11 


354 61 


9 1.5 


57 


10 


6 1 


92 


16 


60 


517 39 


503 38 


24 2 


128 


10 


11 1 


135 


10 


90 


536 38 


522 37 


29 2 


128 


9 


18 1.5 


170 


12 








FIGURE 4 (AREA 4) 










TIME 










% 


-IRE-CRACKED 






ELAPSED 


%LIMESTONE 


% CHERT 


% 


POTTERY 




ROCK % BONE/SHELL 


% MISC 


30 


11 1 1 


61 I 


1 1 


5 


10 


CD 1 


i 


16 CD 


60 


39 i 


38 i 


1 2 


D 



10 
9 


CD 1 
CD 1.5 


i 
a 


10 C3 













Table 5. {Area 5) 



time 












fire-cracked 






elapsed 


limestone 


chert 


pottery 


rock bone/shell 


miscellaneous 




g. % 


g- 


% 


g- 


% 


g- 


% g. 


% 


g. % 


30 


28 3 


652 


69 


28 


3 


220 


23 6 


.5 


14 1.5 


60 


28 2.5 


758 


67 


50 


4.5 


255 


22.5 6 


.5 


35 3 


90 


34 2.5 


836 


59.5 


71 


5 


397 


28.5 6 


.5 


57 4 


120 


34 2.5 


851 


57 


77 


5 


461 


31 11 


.5 


62 4 










FIGURE 


5 (AREA 5) 








TIME 
ELAPSED 

30 


% LIMESTONE 
3 D 

2.5 o 
2.5 D 
2.5 o 


% CHERT 






% 

3 

4. 
5 
5 


POTTERY 


% FIRE-CRACKED 
ROCK 

22.5 i . 


% BONE/SHELL % MISC. 








5 D 

□ 
a 


.5 
.5 
.5 




60 








i 3 a 
















] 4 a 

1 4 D 


120 


57 i 






31 i 1 



It is the conclusion of this study that if the conditions are properly controlled, 
and if a knowledge of the gross classes of material present on the site is the goal 
of an intensive site survey, then the first 30 minutes spent in each grid unit dur- 
ing an intensive surface survey will produce essentially the same set of data as 
does a total surface pick up. It also should be mentioned here that the preliminary 
analysis conducted on the recovered material indicates that over 90% of all 
diagnostic pieces (e.g. rim sherds, decorated sherds and projectile points) were 
collected in the first 30 minute period. 

Finally, it is concluded that the use of this procedure will save substantial 
amounts of time in situations requiring either an intensive survey of a total site 
surface or in situations calling for an intensive survey of a systematically chosen 
sample of a site surface. In the present experiment the five 6M x 6M test units re- 
quired 2.5 man hours for a time-controlled pick up and 16 man hours for a total 
surface pick up. The time-controlled method yielded the same results with an 
84% savings in the required man hours. If one were to generalize this figure to 



Anthropology 79 

the thousands of square meters covered annually with intensive surveys, the 
potential savings in man hours are staggering. It is the hope of the present author 
that these potential savings will merit further experiment and refinement of this 
method by other archaeologists in the field. 



The Fate of the Soul in Modern Aztec Religious Thought 

Paul Jean Provost 

Department of Sociology and Anthropology 

Indiana University-Purdue University at Fort Wayne, Fort Wayne, 

Indiana 46805. 

Introduction 

In this report I would like to describe an incident that occurred during my 
anthropological fieldwork among the Nahua Indians of the Huasteca region of 
northern Veracruz, Mexico. The incident is a tragic one — the funeral of a young 
girl, age twelve, who died one day by drowning during an outing with her 
playmates. The girl was my goddaughter; and because of this, I was asked to spon- 
sor her funeral ritual. The incident was obviously a sorrowful occasion, but it was 
also a rare and unique experience that provided me with a glimpse of the complex- 
ity of Nahua cosmological and eschatological beliefs. 

Sir Edward B. Tylor has written that the essence of any religion is the belief 
in souls (6). In fact, according to Tylor, the heart of religion per se is the concept of 
the soul. That is, if we define religion as the concern with the supernatural, 
spiritual or sacred, then ideas concerning the nature of the soul are the core of 
any belief system we label as religion. Therefore, the best way to understand 
another religious system is to first understand its assumptions concerning the 
nature and fate of the soul. 

Background and Cosmology 

The Huasteca region of northern Veracruz is a hilly, physically remote 
tropical jungle region that lies in eastern Mexico between the Tropic of Cancer 
and the 20th parallel. The area is inhabited by four linguistically distinct Indian 
groups: the Huastecs, the Tepehuas, the Otomi, and the Nahus. By far the most 
numerous of all these groups are the Nahuas, the contemporary descendants of 
the ancient Aztecs who settled in this region in the sixteenth century. 

Like their neighbors, the modern Aztecs are of basically self-sufficient 
subsistence, slash and burn horticulturists, and can be characterized generally 
as falling near the folk end of the folk-urban continuum. That is, they are an 
unacculturated group living in a remote area, following a way of life that allows 
them to maintain a high degree of cultural identity as Indians. Their farming 
equipment consists primarily of the simple digging stick and the machete. Maize 
is the principal crop which is supplemented by black beans, squash, tomatoes, 
chile peppers and various tropical fruits and vegetables. Animal husbandry is 
basically limited to pigs, turkeys and chickens. 

The residential pattern among the modern Aztec is virilocal. That is, 
although each nuclear family has its own dwelling which is built by the male just 
before marriage, the new home is located close to the home of the male's father. 
However, the new dwelling is seen as a separate entity and is built usually at a 
small distance from the home of the male's natal family and connected to it by a 
trail. Thus, the net result of this settlement pattern is a loose grouping of huts 
clustered in one particular area but with each having its own sphere of existence 
separate from that of others. The village then is like a series of dispersed clusters 
of huts or compounds occupied by related males and their families, and connected 
to each other by a labyrinth of jungle trails. 

80 



Anthropology 81 

The cosmology of the modern Aztec is even more complex than their cultural 
geography. For the modern Aztec, like many other peoples, the universe is divided 
into several realms, each inhabited by different spiritual beings. However, these 
are not exclusive and mobility is possible between the realms, a typical situation 
in many of the world's cosmologies. For example, in Greek mythology Zeus, the 
king of the gods, lived on Mt. Olympus, his brother Poseidon lived at the bottom 
of the ocean, while another brother, Pluto, lived under the earth. Nonetheless, 
visitations, invasions or movement between all of these worlds were possible, 
even probable. So too for the modern Aztec. The first realm of the cosmos is 
Ilwihkaktli, the arc of the Heavens. Similar to Greek cosmology this realm is 
home to the most important of the supernaturals that inhabit the Aztec universe. 
Here lives Tonantsi mother of all life and controller of all fertility and reproduc- 
tion. She is associated with a sacred "court" that consists of many vague entities 
syncretized with various figures drawn from the list of Roman Catholic saints. 
Tonantsi also has four sons who are felt to be arranged in importance according to 
age. The oldest and the most important son is Tlakatekolotl the Dark Lord of the 
Underworld. More like Pluto than our Devil, Tlakatekolotl is an ambivant figure 
who rules over Miktlan, a kingdom of ghosts, shades, and spirit-helpers called 
mecos. The second son of Tonantsi is St. John the Baptist and he is married to 
Apanchane, the Lady of Water. The connubial couple control the realm of water 
including the rains, springs, rivers and streams. The water world, called Apan, is 
truly a fluid one and is felt to connect all four levels of the cosmos into an inte- 
grated whole. Rain falls from the sky, while streams, rivers and springs flow on 
the surface of the earth. Also, the modern Aztecs believe that springs emerge 
from and rivers return to the Underworld. Streams and rivers also give water 
back to the sky through evaporation. 

Tonantsi 's third son, Moctezoma, lives on earth in stone ruins that dot the 
surrounding jungle. In reality these stone ruins are ancient pre-Columbian 
pyramids and the accuracy of this connection, which is historically unknown to 
these people, is a testimony to the observation made by French anthropologist 
Claude Levi-Strauss that cultures retain in their ideational systems accurate 
structural and historical data (1). Moctezoma is associated with dark powers and is 
felt to be related to a class of creatures known as Ehecatl or winds that bring 
illness and misfortune (4). 

Tonantsi has one more son, Jesus Christ. He is the youngest son and is 
sometimes syncretized with the sun in the sky (3). For the Aztecs, Christ is 
associated with the saints of Tonantsi 's court; sometimes he is spoken of as the 
"King of Saints" but by and large he is a relatively minor figure whose specific 
qualities are quite vague and generalized. His presence in the modern Aztec pan- 
theon seems to represent the general process of syncretism common throughout 
Mexico at large. It is certain that the figure of Christ was added to the folk Aztec 
pantheon after the general introduction of Roman Catholicism which followed the 
Spanish conquest of 1521. From the historical point of view, Christ truly is the 
youngest or newest diety in Indian or folk cosmologies found throughout the 
Republic of Mexico. Once again, the Aztec view of the cosmos in a certain way 
actually replicates historical reality. 

Death and the Soul 

On September 13, 1977, Reina Hernandez, age twelve, went swimming with 
her little playmates. It was a typically hot September day and the band of little 
girls were excited about the prospect of a pleasant hour or two playing in the cool 



82 Indiana Academy of Science 

waters of a local spring that flowed into a nearby stream. No one knows how but 
somehow Reina was caught in some weeds and drowned before her playmates 
could help her. There is no way to accurately describe the tragedy of a small girl's 
death. To her parents and friends she was a bundle of joy, happiness and love, and 
for those reasons alone her death was a terrible blow. But, for other reasons also, 
her death was painful. For in a few brief seconds Reina went from being a lovable 
child, the favorite of her family, to a dangerous, marauding and fearful spirit entity 
that for the safety of the family had to be driven away. 

In Aztec cosmology, the souls of those who die naturally go to a spiritual 
underworld called Miktlan. Miktlan is much like our own world except that it is in- 
habited by souls who live a ghostly version of life on earth. They are shade-like 
creatures who lack personalities, and may even evaporate after a few years. 
However, people who have an unnatural death caused for example by drowning, 
murder, lightning, childbirth, war or any violence are felt to become highly charged, 
ambivalent, angry forces. Instead of going to Miktlan these creatures go to Apart. 
the watery realm and can therefore wander, like the water itself, through the 
other realms of the Cosmos. They are extremely unpredictable and dangerous due 
to their anger at the premature loss of their lives. Thus, Reina's death was a dou- 
ble tragedy to her family and her funeral came to have a special importance — it 
had to bury the body and at the same time drive off the potentially harmful soul. 
A difficult combination of emotional needs, to say the least. 

The funeral was held on the day of death — a requisite of any tropical environ- 
ment. The child's body was brought to the family's house and a coffin was imme- 
diately constructed on the spot by a carpenter who had been summoned for this 
purpose. Before the body was placed into the coffin, chalk was sprinkled in the cof- 
fin in the sign of a cross. Then a set of clean clothes that had belonged to the girl 
in her lifetime were laid over the chalk, along with a small pillow for her head. 
The body was placed over this and then covered with a shroud. Further objects 
were now placed into the coffin: a set of new earrings, hair combs, a needle and 
thread, money, as well as a plate and cup. The coffin was then filled with flowers 
and sealed. Incense was lit and the women of the family who had gathered at the 
house covered their faces and broke into a grief-stricken ritual wail that was terri- 
ble to hear. At this, the pallbearers picked up the coffin and started to carry it 
outside; whereupon the girl's mother placed the child's drinking glass, cup and 
plate on the threshold. As the pallbearers stepped over the threshold, they 
deliberately stepped on and smashed the utensils. As this was transpiring, the 
girl's grandmother strangled a white chicken and hung it over the door which was 
then sealed shut for good. The funeral entourage followed the casket to the 
cemetery and there the girl was buried. This ended the first part of the funeral 
ritual. 

The second and most important part of Reina's funeral occurred one week 
after her death. Traditionally, a memorial service is held at this time to celebrate 
the arrival of the soul in Miktlan. However, in Reina's case her soul was still at 
large and therefore her memorial service became an extremely important event. 
Accordingly, it was an elaborate event which began in the afternoon of September 
20, 1977, and continued until dawn of the following day. Musicians were hired to 
provide sacred music throughout the night. Quantities of special foods and drinks 
were prepared and a special altar built and decorated with leaves and flowers. On 
the altar were placed Reina's school books, her remaining clothes and quantities of 
cookies, breads and fruits. Against this altar was laid a huge cross decorated with 



Anthropology 83 

hibiscus flowers. In front of the altar a specially built miniature table was set up 
and surrounded by four posts set in the ground, each topped by a burning candle. 
Under this table were several incense braziers. The table was covered with a pall 
on top of which were placed two dresses and two hair ribbons. On top of this 
another more elaborate cross was laid. This was decorated with a colored paper 
cutting in the form of a rebozo - a typical women's shawl. It represented the dead 
girl. In front of this plates of prepared food were placed along with many baskets 
of flowers. The men sat on one side of the room, the women stood on the other 
side. The musicians played constantly while the women wept and wailed 
throughout the night-long vigil. 

During the wake it is expected that some close relative delivers a eulogy 
about the deceased. In this case it was the girl's father. However, because of the 
nature of the girl's death, his eulogy took on an unusual character. Typical of the 
general pattern he spoke to the dead girl but, interestingly enough, he told her 
again and again she was dead! "You are dead, you are gone, you are no longer, you 
are gone, you are dead," he repeated over and over for thirty minutes, during 
which time the women's ritual wail reached almost hysterical levels. As the wail 
increased and decreased through the night, it became obvious that it did so 
according to some scale only part of which was sorrow, the other part owing to 
the needs of the ritual itself. That is, formal wailing is part of a proper funeral 
ritual and women seemed to be able to begin it as well as end it instantly. One 
minute a woman could be wailing, the next talking to someone or taking care of 
some necessary detail. The morning after the wake the crosses and funeral 
thanatophernalia are carried first to the place of death then later to the cemetery. 
The elaborately decorated cross, symbolizing the girl, is left at the actual place of 
death and told again that the girl is dead. The second cross is left at her grave in 
the cemetery and likewise reminded by speech and wail that the young girl is 
dead. 

Aztec Eschatology 

Monica Wilson has written that, 

Rituals reveal values at their deepest level . . . men express in ritual 
what moves them most, and since the form of expression is conven- 
tionalized and obligatory, it is the values of the group that are revealed 
. . . the study of rituals [is] the key to an understanding of the essential 
constitution of human societies (7). 

To this I would add that analysis of ritual behavior also provides insight into com- 
plex and abstract ideas found in a people's cosmology. For example, the study of 
the funeral ritual for Reina Hernandez reveals some essential elements in the 
eschatology of the modern Aztec. First and foremost the nature of the afterlife 
itself is construed as being not immediately or personally relevant to the lives of 
human beings. The afterlife is primarily a place for supernaturals and not a place 
for individual human continuation. The human connection to the afterlife is, at 
best, minimal or imprecisely conceived. In fact, there seems to be no indication 
that the human soul is an integral part of the spiritual world in any fundamental 
sense at all. In this context it is also interesting to note that for the modern 
Aztecs the soul is not a personalized concept as it is for, say, Christians, who 
assume that the soul is an invisible extension of the human personality capable of 
survival after the death of its corporeal frame. In the eschatological thought of the 
modern Aztec, the soul could be likened to the electricity that illuminates a light 



84 Indiana Academy of Science 

bulb — a separate entity not to be confused with the light bulb itself. When the 
light bulb is burned out the electricity continues and even may be harmful if not 
handled properly. The nature of the soul for the Aztec, therefore, is not imma- 
nent, not inherent, but rather a separate independent and non-personalized force 
that is a necessary condition for life but not synonymous with the individuality of 
the human personally. In this sense, the nature of the Aztec soul could be likened 
to Robert Marett's concept of mana, an impersonal spiritual life-force associated 
with living and sometimes even inert things (2). Likewise, in modern Aztec 
religious thought, the soul is believed to be an impersonal life force, analogous, for 
example, to electricity and not a symbolic expression of the human personality. 

As a logical extension of these beliefs, the modern Aztecs see no relationship 
between morality and the supernatural order at all. Therefore, the fate of the soul 
is not dependent, as it is, say, for Christians, upon the moral or ethical behavior of 
its corporeal counterpart. The problem with the fate of Reina's soul was not 
related to her moral conduct but rather to the unusual nature of her demise. Fear 
of retribution in the afterlife for immoral or unethical behavior on earth is not 
found among the modern Aztecs. In fact, a modern Aztec would feel the Christian 
missionaries' plea to be allowed to "save his soul" to be quite ghoulish and 
perverse. Only witches try to save, steal, or otherwise collect souls and their 
intentions are felt to be basically malevolent. 

In summary, then, Aztec cosmology holds that the nature of the afterlife is 
essentially a non-human or supernatural reality that does not involve humans in 
any organic way. The nature of the soul reflects this belief; it is impersonal and 
separate from the human body and personality. It is a life-force and only accom- 
panies the body during its days of life and is not an extension of it. Finally, the 
fate of the soul has no direct connection to the moral or ethical life of its human 
counterpart. 

For the modern Aztec the world is a wonderful and fantastic but knowable 
reality. The Aztecs' attempt to order, understand and explain the world involves 
a complex and unique cosmological schema that while agreeing with all the other 
world's religions in assuming the existence of two worlds, one natural and the 
other supernatural, it contributes its own specific philosophical interpretation of 
these worlds and their relation to and meaning for human existence. 

While it is beyond the scope of this paper to delve into the actual origins and 
functions of modern Aztec religious thought, certain brief observations can be 
made that at least provide a clue to that problem. Following Durkheim, Guy 
Swanson, in his seminal work, The Birth of the Gods (5) suggests that the 
organization of religious systems of thought rests upon empirical sociological con- 
siderations. Religion, according to Swanson, will adapt social groups to their social 
and biological environments by making "the necessary seem obligatory." In his 
understanding "the necessary" refers to the social, technological, and ecological 
requirements of social groups with the implication being that different levels of 
socio-cultural integration need different types of supporting or compatible 
religious systems. While his data is primarily suggestive, his predictions for the 
nature of a cosmology associated with an atomistic, self-sufficient, horticultural 
society closely parallel the above description of modern Aztec religious beliefs. 
Thus, perhaps, the origins and maintenance of such beliefs represent not so much 
the erroneous thinking of "savages" nor even the philosophical ponderings of 
some brilliant Don Juan; but rather derive from and have meaning for specific 



Anthropology 85 

sociocultural conditions and are designed to adapt human groups to the re- 
quirements of their particular techno-environmental situation. 

The essential point then is that the Aztec system of cosmological thought 
described above may originate in material conditions and function to adapt the 
modern Aztec to the realities of their world. Accordingly, then, modern Aztec 
cosmology may represent not so much a supernatural view of the world but rather 
a super-organic realization of the actual conditions of their existence. Being horti- 
cultural^ self-sufficient they are likewise spiritually self-sufficient. Lacking any 
corporate, hierarchical social structures, they likewise do not recognize any high 
or omnipotent gods who concern themselves with human affairs. And lastly, the 
fate of the soul like the nature of the afterlife itself does not depend in any funda- 
mental way upon human activities. Just as the fate of the individual in life is a 
product of his self-sufficiency, so too is the fate of the soul self-sufficient: the con- 
ditions of earthly existence are therefore projected onto the scene of the 
transcendental. To quote Swanson, "Our understanding of behavior suggests that 
all ideas arise from man's experience with his surroundings" (5). Aztec cosmology 
then may be said to actually replicate Aztec reality. 

Literature Cited 

1. LEVI STRAUSS. CLAUDE. Anthropologic Structural. Paris: Plon, 1958. 

2. MARETT. ROBERT R. Anthropology. New York: Holt Company, 1912. 

3. REYES GARCIA, LUIS. Pasion y Muerte del Crista Sol. Veracruz: Xalapa, 1960. 

4. SANDSTROM, ALAN. The Image of Disease: Medical Practices of Nahua Indians of the 
Huasteca. Columbia, Missouri, 1978. 

5. SWANSON. GUY. The Birth of the Gods: The Origin of Primitive Beliefs. Ann Arbor, 
Michigan, 1968. 

6. TYLOR. SIR EDWARD B. Primitive Culture, Vol. I. New York: Holt Company, 1877. 

7. WILSON, MONICA. "Nyakyusa Ritual and Symbolism," in America Anthropologist, Vol. 
56, No. 2, 1954. 



BOTANY 



Chairman: Gary Dolph 
Department of Botany, Indiana University — Kokomo, Kokomo, Indiana 46901 

Chairman-Elect: Charles T. Hammond 
Department of Biology, St. Meinrad College, St. Meinrad, Indiana 47577 



ABSTRACTS 

A Proposed Third Function for Root Caps. William W. Bloom, Department of 

Biology, Valparaiso University, Valparaiso, Indiana 46383. Root caps are 

believed to protect the delicate meristematic region of the root tip as it is pushed 
through the soil. Evidence suggests that root caps also control positive geotropism. 
No satisfactory explanation exists for the delay in the production of root hairs until 
after the primary root tissues have elongated. Root hairs, like rhizoids of ferns, only 
form on the exposed surfaces of cells. An examination of a number of longitudinal sec- 
tions of root tips indicates that most root caps extend well into the region of elonga- 
tion and closely adhere to the root in this region. The outer surface of the epidermal 
cells are not freed of the root cap until cell elongation has pushed them free of the 
enclosing cap, thus permitting root hair formation. This hypothesis is being tested 
experimentally. 

Utilization of an Artifical Beach in Investigation of Phytopsammon Communities. 

William Da vies, Department of Biology, Indiana University-Purdue University at 

Fort Wayne, Fort Wayne, Indiana 46805. Interstitial voids in freshwater sandy 

beaches harbor well-defined algal communities (phytopsammon). Coefficient of com- 
munity and index of affinity data indicate the phytopsammon communities differ in 
composition from phytoplankton communities of adjacent lakes. Indices of diversity 
for phytopsammon communities along beach transects indicate significant relation- 
ships between phytopsammon community structure and substrate water content, 
substrate grain size, and grain size uniformity coefficients. Chlorophyll extraction 
and 14 C incubation techniques are being utilized in current studies of an artificial 
beach constructed at the Indiana University-Purdue University at Fort Wayne 
biological station at Crooked Lake. The artificial beach has been constructed to 
reduce variables in substrate grain size, grain size uniformity coefficients, detritus 
content and beach substructure. 

A Developmental Study of Foliar Epidermal Features in Castanea mollissima 
Blume (Fagaceae). Gina Fernandez, Department of Biology, Ripon College, Ripon, 
Wisconsin 54971, and Jay H. Jones and David L. Dilcher, Department of Biology, 

Indiana University, Bloomington, Indiana 47405. Leaves of various ages were 

collected from Chinese chestnut trees (C. mollissima) in order to assess changes in 
epidermal features that occur during development. Macerated cuticles were 
prepared for examination with the light microscope by standard methods. Leaves or 
leaf sections were critical point dried and prepared for examination with the scan- 
ning electron microscope. Isolated cuticles were extremely difficult to prepare from 
very young leaves and cleared leaves were used to assess epidermal features in 
early stages of development. Trichome initiation occurs very early in development. 
The highest trichome densities were found in very young ( < 2 cm) leaves. Trichome 

86 



Botany 87 

density declined with leaf expansion on both adaxial and abaxial surfaces. The 
lowest densities occurred in old fully expanded leaves in which many trichomes had 
been lost. The relative proportions of the five trichome types found on these leaves 
varied with age. In addition the distribution of trichomes was also found to be type 
specific. Nonvenous areas tend to lose their pubescence sooner than venous areas. 
The stomatal complex is anomocytic and the development appears to be 
aperigenous. The epidermal development of C. mollissima appears to be represen- 
tative of other Fagaceae. The results of this study provide some basis for determin- 
ing the maturity of fossil Fagaceous leaves. 

Effects of Irradiance on the Morphological Characteristics of Two Plant Species 
of the Maritime Strand. Stephen W. Fletcher, Environmental Science and 

Engineering, Inc., Gainesville, Florida 32604. Previous authors have described 

three types of shade response in plants, including shade tolerance by metabolic 
and morphological adaptation, shade avoidance through stem elongation, and in- 
tolerance with no ability to elongate. In order to examine the ecological implica- 
tions of such responses, the morphological adaptation patterns as a function of 
growth irradiance were studied in Cakile harperi (sea rocket) and Euphorbia 
polygonifolia (seaside spurge), two annual plant species restricted to coastal 
beaches and sand dunes. 

Both species were grown under controlled environments at a common 12-14 h 
photoperiod and three growth temperatures (32/29, 26/23, and 17/11 °C). Mor- 
phological aspects of shade adaptation at irradiance levels from 30-120 W m" 2 
were described by measurements of leaf weight and area per plant, leaf area- 
weight relation, shoot/root ratio, stomatal density, and chlorophyll content. Stem 
length, internode length, and number of internodes were treated separately as 
measures of shade avoidance potential. 

On the basis of field and laboratory survival patterns, chlorophyll levels and 
chlorophyll a/b ratios, both species were determined to be of the shade-intolerant 
type. However, both were found to be capable of some degree of morphological 
acclimation in response to shading. Relative leaf area was higher and stomatal 
density was lower in shade-grown plants. In Cakile harperi the increased relative 
leaf area was due to a decrease in leaf thickness; in Euphorbia polygonifolia this 
was effected by a shift in photosynthate distribution from root to shoot. Both 
species showed the minimum amount of stem elongation necessary to overtop the 
canopy level of dominant dune grass. This avoidance response was dependent 
upon internodal elongation rather than on an increase in the number of elongation 
sites, and upon the resultant stability of the stems. Since the response patterns of 
these species were found to be substantially affected by growth temperature, it is 
recommended that light acclimation studies be conducted as closely as possible to 
the optimal growth temperature or the known growing conditions for each 
species. 

The Ultrastructure of Nonarticulated Branched Laticifers in Asclepias tuberosa 
L. (Butterfly Weed). Vonda Frantz and Kathryn Wilson. Department of 
Biology, Indiana University-Purdue University at Indianapolis, Indianapolis, In- 
diana 46205. Meristem, leaf, and stem tissues from Asclepias tuberosa seed- 
lings and one year old plants were prepared for electron microscopy. Material was 
fixed in Karnovsky fixative (pH 7.0), post-fixed in 1.0 percent osmium tetroxide, 
dehydrated to acetone, and embedded in Spurr low-viscosity resin. Embedded 
material was sectioned, mounted on uncoated 300 mesh copper grids, stained in 
lead citrate and uranylacetate, and examined on an RCA EMU-3G electron 



88 Indiana Academy of Science 

microscope at 50 KV. The ultrastructure of laticifers in Asclepias tuberosa was 
found to be comparable to nonarticulated branched laticifers in other Asclepias 
species. Maturing laticifers of A. tuberosa are characterized by an extensive 
vacuolar system, apparently degenerating plastids, an absence of plasmodesmata 
in the cell walls, and by their long branching morphology. A large central vacuole 
appears to develop by dilation of endoplasmic reticulum and by coalescence of 
small vacuoles in the maturing cytoplasm. Mature laticifers differ markedly for 
those of A. syriaca and A. curassavica because the large central vacuole does not 
contain rubber particles or other electron dense secretory products. 

Paleobotanical Nomenclature: Principles, Problems, and Proposals. Jay H. Jones 
and David L. Dilcher, Department of Biology, Indiana University, Bloomington, 
Indiana 47405. The naming of fossil plants has posed problems since the incep- 
tion of "modern" binomial nomenclature. The fragmentary nature of fossil re- 
mains and the fact that they cannot be interbred to test the validity of biological 
species prevent easy application of all rules in the International Code of Botanical 
Nomenclature. Attempts to modify the Code to accomodate fossil and other excep- 
tional forms have historically led to the formation of special taxa, i.e. organ genera 
and form genera. Under the current code, fossil genera are functionally equivalent 
to extant genera; and the former taxa have been eliminated. There is still con- 
siderable disagreement as to whether or not these special taxa should be used. In 
addition to these disputes paleobotanists also concern themselves with the vali- 
dity of applying modern generic or specific names to fossil materials. A somewhat 
related philosophical problem is whether or not to indicate the suspected tax- 
onomic affinities of a fossil in its name. Such names can be quite misleading when 
the originally suspected affinities are proven incorrect and the original name must 
be conserved. The above problems are substantially a function of one's personal 
philosophy of how to deal with problems unique to fossils. A scan of the 
paleobotanical literature, however, quickly reveals frequent and flagrant 
disregard of parts of the ICBN for which fossils pose no particular problem. The 
localized trouble spots in the Code seem to lead some paleobotanists to loosely in- 
terpret the whole Code. A thorough reexamination of the purpose and practice of 
naming fossils seems to be in order, so that a precise and functional system can be 
established. 

Fruits of the Pterocarya alliance (Juglandaceae) from the Paleogene of the Rocky 
Mountain Region. Steven R. Manchester and David L. Dilcher, Department of 

Biology, Indiana University, Bloomington, Indiana 47405. Paleocene and 

Eocene deposits of the Rocky Mountain region have yielded three genera of 
winged fruits belonging to the Pterocarya alliance of the walnut family. Two of 
the genera are present in the modern flora of Asia, while the third represents an 
extinct form. Two fossil species from the Rocky Mountain region belong to the 
extant genus Pterocarya. Both are bi-winged, as are modern species of the genus. 
One of the species, P. roanensis Mac Ginitie, occurs in the Eocene Green River 
flora of Utah. Another species, with narrower wings, occurs in the as yet 
undescribed Eocene Wind River flora of Wyoming. The extant pterocaryoid genus 
Cyclocarya is represented by the species which Brown formerly called Pterocarya 
hispida, from the Paleocene Fort Union Formation. New collections of this species 
from North Dakota indicate that, like the living species Cyclocarya paliurus, the 
fossil had a seed which was 4-lobed at the base, surrounded equatorially by a 
single, large flange-like wing. A new extinct genus from the Paleocene Fort Union 
flora was previously described by Brown (1962) as Pterocarya glabra. This genus 



Botany 89 

differs from modern Pterocarya and Cyclocarya in having numerous (8 to 10) 
wings. The wings are arranged equatorially about a 4-lobed seed. Although it is 
pterocaryoid in its affinities, it can not be retained in the extant genus 
Pterocarya. In the present day flora there are only two genera in the Pterocarya 
alliance: Pterocarya and Cyclocarya. However, as indicated above, there were at 
least three genera in the Paleogene. This illustrates a pattern of decreasing 
generic diversity similar to that observed in the Engelhardia alliance of the same 
family. Some of the genera present in the early Tertiary, such as Pterocarya and 
Cyclocarya, have persisted to the present day while others, such as the multi- 
winged form from the Fort Union Formation, have since become extinct. 

Stomatal Development in Asimina triloba |L.) Dunal. John L. Roth, Jr. and David 
L. DlLCHER, Department of Biology, Indiana University, Bloomington, Indiana 
47405, and Brent A. McKim, North Harrison High School, Ramsey, Indiana 47166. 

Asimina triloba (L.) Dunal, commonly known as the Pawpaw, grows in 

temperate forests of eastern North America from southern Canada to the Gulf 
Coast. As such it is unique among the more than 2000 species of the tropical family 
Annonaceae. This provides an opportunity to assess the effects of environment on 
stomatal ontogeny by comparing stomatal development of the temperate and 
tropical species. The stomatal complex of Asimina triloba is generally 
brachyparacytic, but paracytic, hemiamphibrachyparacytic and amphibrachypar- 
acytic variations are common. Its stomatal development is syndetocheilic. A rec- 
tangular meristemoid divides to form two cells of unequal size. The smaller of the 
two divides into two narrow cells forming a complex of three cells. The center cell 
divides to form the guard cells which develop an intervening pore. The two cells 
bordering the guard cells are the subsidiary cells and may be non-contiguous around 
the ends of the guard cells or contiguous at one or both poles. They enlarge and may 
undergo an additional division to form three or four subsidiary cells. Although 
Asimina triloba has been subjected to the rigors of a temperate environment since 
at least the Pleistocene, its stomatal development has remained essentially un- 
changed from that reported for its tropical relatives. This suggests that stomatal 
development is under rather strict genetic control and is not easily modified by en- 
vironmental stress. 

Bisexuality in the Pistilate Inflorescence of Plat anus Occident alis. L. Robert 
Schwarzwalder, Jr. and David L. Dilcher, Department of Biology, Indiana 

University, Bloomington, Indiana 47405. Platanus occidentalis L., the plane 

tree or sycamore, is a common plant along streams and rivers over much of Eastern 
North America. Consisting of seven to eleven species, the monotypic family 
Platanaceae is generally considered to have monoecious trees with unisexual in- 
florescences. Dissections of pistilate inflorescences of Platanus occidentalis reveal 
tight clusters of floral units each of which typically has nine carpels, three or four 
staminodia and an encircling membranous sheath. The staminodia commonly are 
assumed to be phylogenetically reduced stamens and the membranous sheath is 
hypothesized by some to represent a vestigial perianth. In examining numerous 
pistilate inflorescences we found varying degrees of bisexual development ranging 
from, rarely, fully functional stamens and carpels to the more common condition of 
sterile staminodia surrounding functional carpels. Pollen from stamens obtained 
from pistilate inflorescences was tested and found to be viable. While infrequent in 
Platanus occidentalis, bisexuality raises several phylogenetic questions. Whether 
this condition is an expression of the potential for bisexual development in Platanus 
or represents a reduction from a previously bisexual flower is difficult to ascertain 



90 Indiana Academy of Science 

without paleobotanical evidence. Our observations also raise questions concerning 
the sexual plasticity that may be expressed in flowering plants. While often assum- 
ed to be a static morphological feature, the formation of unisexual inflorescences in a 
monoecious species could be interpreted as one of a range of responses to a myriad of 
environmental pressures. 

Endomycorrhizae Increases Growth of Sycamore Seedlings. James P. Shepard 
and William R. Chaney, Department of Forestry and Natural Resources, Purdue 

University, West Lafayette, Indiana 47907. Sycamore {Platanus occidentalis 

L.) germinants were transplanted into 45 six-inch-pots containng low phosphorus 
(1 ppm), sandy loam soil mix. Spores of the endomycorrhizal fungus Glomus 
fasciculatus (menge isolate) were incorporated as pot culture inoculum into soil of 
one-third of the pots at a rate equivalent to 2000 spores per ft 2 surface area. An 
equal amount of inoculum was filtered through a 44 (im sieve to remove fungal 
spores and the filtrate added to another one-third of the pots to equilibrate other 
organisms and compounds present in the inoculum. One-third of the pots were un- 
treated. Seedlings were grown in a greenhouse for 20 weeks. Growth was quan- 
tified at four-day intervals using the plastochron index, an estimate of mor- 
phological rather than chronological age. Inoculated seedlings had a greater (p < 
.01) plastochron age than filtrate treated and control seedlings. Regressions of 
plastochron index with time revealed a higher rate of leaf initiation by inoculated 
than non-inoculated seedlings. Endomycorrhizae inoculated seedlings also had a 
greater (p < .01) height and stem diameter and leaf and stem dry weight than un- 
treated seedlings after 20 weeks of growth. The inoculum filtrate did not 
significantly influence seedling growth. 

The Distribution and Uses of Arundo donax. Marilyn Sue Veselack and Jerry J. 

Nisbet, Ball State University, Muncie, Indiana 47306. Arundo donax is widely 

distributed in warm sunny climates and is currently found growing in India, 
Burma, China, Southern Africa, Australia, in the Americas, in regions adjoining 
the Nile River and in the Mediterrean area. Historically, Arundo donax has been 
used by man for a variety of purposes. Starting as early as 5,000 B.C. Egyptians 
used Arundo to line underground pits used for grain storage. Mummies of the 
fourth century A.D. were wrapped with Arundo, and the plant has been adapted 
to a variety of fabrication uses including the making of fences, awnings, baskets, 
fishing nets, writing pens, and fishing poles. Pulp from Arundo has been used in 
making paper and rayon. A number of extracts with medicinal properties have 
been derived from the plant including donaxine, bufotenidine and dehydrobufo- 
tenine. Arundo donax has been significant to many civilizations for its use in the 
construction of musical instruments and in the manufacture of reeds for musical 
instruments. No synthetic reed material has as yet been manufactured which can 
match the tone quality contributed to woodwind instruments by high quality 
Arundo reeds. 

Squirrel Resistant Black Walnuts for Direct Seeding? Not Yet. Robert D. 

Williams, U. S. Forest Service, Bedford, Indiana 47421. We have observed in 

some of our nursery studies and direct seeding studies, where seed source was a 
variable, that squirrels seem to prefer the nuts from some trees over those of 
others. In a period from 1975 to 1979, in an effort to find a tree bearing squirrel 
resistant nuts, seeds from 55 trees have been offered to squirrels at 4 locations, in 
fall and spring sown direct seeding trials. We didn't find a mother tree that bears 
squirrel resistant nuts. We did confirm that squirrels prefer the nuts from certain 



Botany 91 

trees; but after the preferred nuts are consumed, they take the less preferred 
nuts. 

Air Toxicity to Eastern White Pines in Indiana. Wayne T. Williams and Roland 

W. Usher, The Institute of Ecology, 4600 Sunset Avenue, Indianapolis, Indiana 

46208. Air pollution disease is widespread on eastern white pine [Pinus 

strobus L.) in Indiana. Ozone concentrations sufficient to elicit phytotoxicity occur 
regionally numerous times each growing season, and sulfur dioxide concentrations 
considered to be phytotoxic are common at urban sites and rural areas downwind 
from coal-fired power plants. Severity of disease on the second needle complement 
differed in 1979 and 1980 in response to the regional air pollution load in 1978 and 
1979. Tip necrosis of needles was more prevalent than needle flecking and 
chlorotic mottling, but both symptom types are considered primary manifesta- 
tions of air pollution disease. Premature needle abscission was found to be 
widespread and more common in areas with more air pollution. But abscission was 
not as closely associated with disease severity as other symptoms including 
chlorosis/flecking, percent of trees with tip necrosis on the majority of foliage, and 
length of necrosis, all of which correlate with disease severity according to linear 
regression analyses. A disease index composed of these four components reflected 
changes in disease severity from year to year, and within and among study sites. 
Changes in frequency distribution of the principal index components are discussed 
in relation to changes in disease severity along a gradient of differing pollution 
concentrations and durations. 



Phosphate Stimulates or Inhibits Silicomolybdate 
Reduction in Spinach Chloroplasts 

Rita Barr and Frederick L. Crane 

Deparment of Biological Sciences 

Purdue University, West Lafayette, Indiana 47907 

Abbreviations used: ADP-adenosine phosphate dinucleotide; DCMU-3-(3,4-dichloro- 
phenyl)-l,l dimethylurea; MV-methylviologen; PS I - Photosystem I; PS II - Photosystem II. 

Introduction 

Silicomolybdic acid as an electron acceptor in chloroplasts was first described 
by Giaquinta et al (7) and Barr et al. (4). Two sites were found in PS II: one, which 
was sensitive to DCMU, another, which was DCMU-insensitive. Berg and Izawa 
(5) showed that the DCMU-insensitive site could phosphorylate with a P/e 2 ratio of 
0.2 in presence of protective agents, although Giaquinta and Dilley (6) had shown 
that no H + were transported in that portion of the electron transport chain from 
the water oxidation site to Q, the primary electron acceptor in PS II. After Barr 
and Crane (3) found 2 DCMU-insensitive silicomolybdate reduction sites in PS II of 
spinach chloroplasts, depending on the pH of the reaction medium, it became man- 
datory to describe all possible differences between these 2 sites. The present 
study was undertaken with the purpose of finding out, how anions affected 
silicomolybdate reduction in PS II. 

Materials and Methods 

Chloroplasts were prepared from market spinach in 0.4 M sucrose and 0.05 M 
NaCl as previously described (12). Chlorophyll was determined according to 
Arnon (1). Oxygen evolution or uptake was measured with a Clark-type electrode 
in illuminated chloroplasts according to Troxel et al. (12). Reaction mixtures for 
assays are given in figure legends. Reaction rates were recorded with a Sargent- 
Welch SRG recorder. 

The anions used in this study were prepared as 0.25 M stock solutions of the 
sodium salts of phosphate, arsenate or sulfate. The pH of phosphate solutions was 
adjusted to 6 or 8 with an Orion pH meter, using mono and divalent phosphate 
salts; the pH of arsenate solutions was adjusted to pH 6 or 8 with HC1. Sulfate 
was used without adjustment of pH, the pH of Na sulfate solutions being « 6.5. 

Results and Discussion 

In order to find differences between the 2 DCMU-insensitive silicomolybdate 
reduction sites first described by Barr and Crane (3), hitherto separated on the 
basis of pH and various inhibitors, the effect of anions on these 2 sites was in- 
vestigated in this study. 

Previous studies, too numerous to discuss in detail here, have shown that 
ions can affect electron transport in chloroplasts in various ways, including a 
direct effect on the light harvesting pigment-protein complex (11), an effect on 
CF X (11), the coupling factor, or an effect on grana stacking (11). Alteration of local 
surface charges by ions (9) also have been investigated. In this study we have ex- 
amined the effect of 3 anions — phosphate, arsenate, and sulfate — on electron 
transport under phosphorylating and under uncoupled conditions, in presence or 
absence of additional buffers. Figures 1A and IB show that electron transport 
through PS I and II in a buffered system is only slightly stimulated by phosphate 

92 



Botany 



93 




IA 



O phosphate (1-IOmM) 

• ADP <0 2-2mM) 

O phosphote dOmM) ♦ AOP !0 2-2mM) 



2 4 6 8 10 

Compound Added 



^ 100 
& 



IB 



phosphote (1- lOmM) 

• AOP (0 2-2mM) 

» phosphote (10mM) + ADP l02-2mM) 



-•-•-* 



2 4 6 8 

Compound Added 



IC 




O phosphote d-10mM) 

• ADP (0 2-2mM) 

8 phosphote dOmM) + AOP (0 2-2mM) 




(1-IOmM) 

-2mM) 
(10mM)» ADP(0 2-2mM) 



2 4 6 8 

Compound Added 

Figure IAD. Stimulation or Inhibition of Electron Transport Reactions in 
Spinach Chloroplasts by Phosphate in a Buffered System. 1-A. H<ft — MV 
(+ azide), pH 8; 1-B H 2 - SM ( + DCMU), pH 6; 1-C H 2 - SM ( + DCMU), pH 
8, and 1-D H 2 — SM, pH 8. The reaction mixture for 1-A contained 25 mM Tris- 
Mes, pH 8, 0.5 mM Na azide and 0.5 mM MV; for IB 0.25 mM Tris-Mes, pH 6, 5 
nM DCMU, and 0.075 mM SM; for 1-C 25 mM Tris-Mes, pH 8, 5 pM DCMU, and 
0.25 mM SM; for ID 0.25 mM Tris-Mes, pH 8, and 0.25 mM SM. Other compounds 
were added to these reaction mixtures as indicated. 



in the H 2 — MV (+ azide) reaction and in the H 2 — SM ( + DCMU) reaction at 
pH 6 in PS II, while phosphate inhibited both the DCMU-sensitive and the DCMU- 
insensitive silicomolybdate reduction at pH 8. This inhibition of electron transport 
by phosphate at pH 8 is not associated with uncoupling, because it is not 
eliminated in uncoupled chloroplasts (Table 1). This effect can also be considered 



94 Indiana Academy of Science 

TABLE I. Stimulation of Silicomolybdic Acid Reduction By Phosphate Anions In Spinach Chloroplasts In 
Absence of Tris-Mes Buffer and NH.Cl and Decreased Stimulation In Buffered, Uncoupled Chloroplasts 



Reaction 


Phosphate cone. 


for 


Stimulation by phosphateDecreased stimulation by 




maximum stimulation 


in unbuffered chloro- 


phosphate 


in buffered, 








plasts 


uncoupled 


chloroplasts 




(mM) 




rate 1 ' 2 ! 


o/o 31 


rate W 


% 3 1 


H 2 - SMUDCMU), pH6 














reaction rate ( - buffer) 


10 




45 


- 


192 


_ 


reaction rate ( + phosphate) 


10 




124 


+ 176 3] 


192 





H 2 - SM( + DCMU),pH8 














reaction rate ( - buffer) 


5 




62 


- 


107 


_ 


reaction rate ( + phosphate) 


5 




118 


+ 90 


85 


-21 


H 2 - SM, pH 8 














reaction rate ( - buffer) 


5 




51 


- 


136 


_ 


reaction rate ( + phosphate) 


5 




146 


+ 186 


79 


-42 


H 2 - MV ( + azide), pH 8 














reaction rate ( - buffer) 


10 




203 


- 


919 


_ 


reaction rate ( + phosphate) 


10 




519 


+ 156 


896 


-2 



1] Reaction rate equals /iequiv./mg chl • hr 

2] The composition of reaction mixtures is given in figure legends 

3] + indicates % stimulation over control rates. 



an anti-uncoupling effect. Its origin is unknown at present. Arsenate (Figures 2A 
-C) showed a comparable slight stimulation of the various reactions and an inhibi- 
tion of silicomolybdate reduction at pH 8. A slight stimulation in place of inhibi- 
tion in the overall silicomolybdate pathway (Figure 2-D) by arsenate also occurred. 
Arsenate had previously been shown to stimulate electron transport through un- 
coupling (8). Sulfate (Figures 3A-D) was not a good stimulator of electron 
transport in this study, which shows that the phosphate and arsenate effects were 
not simple ionic strength effects, compared to phosphate or arsenate. Sulfate was 
shown by Pick and Avron to act as an energy transfer inhibitor (10). Stimulation 
by phosphate in all the reactions studied was greatest, when no other buffer was 
present in the reaction mixtures. Under these conditions (Table 1) electron 
transport rates were stimulated from 90-186% in absence of ADP. 

Stimulation of electron transport by anions appeared to be influnced by un- 
coupling. When NH 4 C1 was present (Table 1), stimulation was abolished, implying 
that phosphate, arsenate, and sulfate could stimulate electron transport by acting 
as uncouplers themselves. This is not surprising, since arsenate has been used as 
an uncoupler in mitochondria. 

The stimulation of electron transport by phosphate in the DCMU-insensitive 
silicomolybdate pathway at pH 6 is consistent with Berg and Izawa's (5) inter- 
pretation, that this short segment of the electron transport chain represents a PS 
II energy conservation site with a P/e 2 of 0.2. 

In summary, this study shows that anions can stimulate overall electron 
transport and the DCMU-insensitivie silicomolybdate reduction at pH 6, while in- 
hibiting all silicomolybdate pathways at pH 8. The order of effectiveness of anions 
is as follows: phosphate > arsenate > sulfate. The mechanism of this stimulation 
may be related to an uncoupling-type reaction, involving the coupling factor, 
although lesser effects by anions, such as alteration of local surface charges, can- 
not be excluded as contributing factors of the stimulation observed. 



Botany 



95 



ha 




O arsenate (1-10mM) 

• ADP (0 2-2mM) 

« arsenote UOmM) ♦ ADP (02-2mM) 



150 r 



HB 



&t**fttt% 



3- 50 



2 4 6 8 10 

Compound Added 



O arsenate d-10mM) 

• ADP <0 2-2mM> 

« arsenote (10mM) ♦ ADP(0 2-2mM) 



2 4 6 8 

Compound Added 



nc 




O arsenate (1-10mM) 

• ADP (02-2mM) 

O orsenate (10mM)+ ADP(02-2mM) 



HD 



2 4 6 

Compound Added 



2-^-^r— cr-rr 

T— Z— *-o-w- 



O arsenate (1-10mM) 

• ADP(02-2mM) 

9 arsenate ClOmM) + ADP(02-2mM) 

_a_a_n—Q-o-c 



cr 
o 

3. 50 h 



2 4 6 8 

Compound Added 



Figure 2A-D. Stimulation or Inhibition of Electron Transport Reactions in 
Spinach Chloroplasts by Arsenate in a Buffered System. 2-A H 2 — MV ( + 
azideh pH 8; 2-B H 2 - SM(+ DCMU), pH 6; 2-C H 2 - SM(+ DCMU), pH 8; 
2D H 2 — SM, pH 8. Reaction mixtures as in Fig. IAD. 



96 



Indiana Academy of Science 



UTA 



O sulfate (1-10mM) 
• ADP (02-2mM) 
9 sulfate (10mM) + ADP (0 2-2mM) 



hib 



O sulfate (1-10mM) 
• ADP(0.2-2mM) 
Q sulfate (10mM) + ADP(02-2mM) 



^Eg^gSM^ 



2 4 6 8 

Compound Added 



2 4 6 8 10 

Compound Added 



mc 



HID 



sulfale (1-10mM) 
ADP (0 2-2mM) 
sulfate (10mM) + ADP (0 2-2mM) 



2 4 6 8 10 

Compound Added 




O sulfate (1-10mM) 
• ADP (02-2mM) 
© sulfate (10mM) + ADP (02-2mM) 



2 4 6 8 10 

Compound Added 



Figure 3A-D. Inhibition of Electron Transport Reactions in Spinach Chloroplasts 
by Sulfate in a Buffered System. 8- A H 2 - MV (+ azide), pH 8; SB H 2 - SM 
(+ DCMUK pH 6; S-C H 2 - SM(+ DCMUK pH 8; 3-D H 2 - SM, pH 8. Reac- 
tion mixtures as in Fig. IAD. 



Botany 97 

Acknowledgements 

This study was supported by N.S.F. Grant # PCM-7820458. 

Literature Cited 

1. Arnon, D. I. 1949. Copper enzyme in isolated chloroplasts. Polyphenoloxi- 
dase in Beta vulgaris. Plant Physiol. 24, 1-15. 

2. Barber, J. 1976. Ionic regulation on intact chloroplasts and its effect on 
primary photosynthetic processes. In The Intact Chloroplast. J. Barber, ed. 
Elsevier/North Holland Biomedical Press. The Netherlands. 476 pages, pp. 
89-134. 

3. Barr, R. and F. L. Crane. 1980. Two possible 3-(3,4-dichlorophenyl)-l, 
1-dimethylurea-insensitive sites in photosystem II of spinach chloroplasts. 
Biochim. Biophys. Acta 591, 127-134. 

4. Barr, R. , F. L. Crane and R. T. Giaquinta. 1975. Dichlorophenylurea-in- 
sensitive reduction of silicomolybdic acid by chloroplast photosystem II. 
Plant Physiol. 55, 460-462. 

5. Berg, S. P. and S. Izawa. 1977. Pathways of silicomolybdate photoreduction 
and the associated photophosphorylation in tobacco chloroplasts. Biochim. 
Biophys. Acta 460, 206-219. 

6. Giaquinta, R. T. and R. A. Dilley. 1975. A partial reaction in photosystem 
II: reduction of silicomolybdate prior to the site of dichlorophenyldimethy- 
lurea inhibition. Biochim. Biophys. Acta 387, 288 305. 

7. Giaquinta, R. T., R. A. Dilley, F. L. Crane and R. Barr. 1974 Photo- 
phosphorylation not coupled to DCMU-insensitive photosystem II oxygen 
evolution. Biochem. Biophys. Res. Communs. 59, 985-991. 

8. GOOD, N. E. 1977. Uncoupling of electron transport from photophosphoryla- 
tion in chloroplasts. In Encyclopedia of Plant Physiology, vol. 5. Photo- 
synthesis I. A. Trebst and M. Avron, eds. Springer-Verlag. Berlin. 730 pages, 
pp. 429-436. 

9. Masamoto, K., S. Itoh and M. Nishimura. 1980. Salt-induced pH changes in 
spinach chloroplast suspension. Changes in surface potential and surface pH 
of thylakoid membranes. Biochim. Biophys. Acta 591, 142-152. 

10. Pick, U. and M. Avron. 1973. Inorganic sulfate and selenate as energy 
transfer inhibitors of photophosporylation. Biochim. Biophys. Acta 325, 
297-303. 

11. Thornber, J. P. 1975. Chlorophyll-proteins: light harvesting and reaction 
center. Ann. Rev. Plant Physiol. 26, 127-158. 

12. Troxel, K. S., R. Barr and F. L. Crane. 1980. The role of Ca 2+ in electron 
transport of spinach chloroplasts. Proceed. Indiana Acad. Sci. 89, 343-349. 



Juglone Dermatitis: Allergy or Irritant? 

Deborah Williams Craton 

Indiana University School of Medicine 

Indianapolis, Indiana 46223 

and 

Robert D. Williams, Principal Silviculturist 

North Central Forest Experiment Station, 

Bedford, Indiana 47421 

Introduction 

Juglone, a powerful chemical (C 10 H 6 O 3 ) found in black walnut {Juglans nigra L.) 
and other Juglans species, is known to be toxic to some neighboring plants, and 
juice from the hulls of the nuts has caused dermatitis on some people hulling the 
nuts. Two U. S. Forest Service employees developed a blistering type rash follow- 
ing exposure to black walnut hulls and bark chips. A case history concerning the 
exposure and reaction of one of the people states that on a hot, humid day in Oc- 
tober he was hulling black walnuts by squashing them between two bricks, thus 
splattering juice from the hulls over his face, hands and forearms. "That night, 
after I had gone to bed, my face and arms started itching. The next morning my 
face, hands and arms were covered with red, itchy bumps that looked like poison 
ivy. Some areas, especially around the wrists, had already begun to blister." 

The other afflicted Forest Service employee, a technician who worked daily 
with black walnut, wrote: "On June 29, I went to the Blackwell tract to prune 
limbs off black walnut trees. I was using a small chain saw. The temperature was 
ranging from 95° to 100° F, with very high humidity. As I pruned limbs that were 
chest high, the wood chips from the saw were sticking to my arms, face and neck. 
At approximately noon, I noticed my arms were stained and starting to burn. By 1 
or 2 p.m., the burning was getting pretty painful. The next morning there was 
some swelling on my arms; several blisters had formed; and some of them had 
burst and were running." 

Because several hundred people work in black walnut plantations or gather 
and hull the fruit, we decided to determine whether the dermatitis caused by 
walnut juice (probably juglone) is an irritant or an allergic reaction. 

Methods and Procedures 

To determine the type reaction we used two methods: questionnaires and 
patch tests. First, we thought it was necessary to determine whether other people 
had experienced similar problems with black walnut. So, in the Walnut Council 
Newsletter of July 1979 Funk and Williams published an article entitled "Can 
Juglone Be Hazardous To Your Health?" In the article a request was made for 
those who had suffered similar experiences with black walnut to notify us. Then a 
questionnaire dealing with the following was sent to the persons responding to 
the article: 

1. Why there was exposure to walnut. 

2. Time of year person was exposed. 

3. Temperature and general weather conditions. 

4. Person's physical condition. 

98 



Botany 99 

5. Parts of the body affected. 

6. Time elapsed between exposure and evidence of reaction. 

7. Similar reactions before or since exposure. 

8. Past history of exposure to black walnut. 

Second, we patch-tested a number of individuals to determine: (1) the type of 
reaction produced by black walnut, (2) the time period required to produce a reac- 
tion, and (3) what part of the black walnut tree actually produced a reaction. 

We first tried a commercially prepared extract from Hollister-Stier 
Laboratories', a wood oleoresin in alcohol, which was a 1:10 strength. The extract 
was placed on a testing patch and taped to the inner forearm for 48 hours. Ten 
subjects, including the two foresters known to have reacted to black walnut, were 
tested. None of the 10 volunteers reacted. Either the strength of the commercial 
extract was not sufficient to cause reaction, and/or the extract did not contain 
some vital material from the black walnut. 

After this failure, we devised a way to use a piece of black walnut bark taped 
to the inner arm during the summers of 1979 and 1980. The 1979 test was done on 
six Hoosier National Forest employees, including the two Forest Service people 
known to be susceptible to walnut dermatitis. Bark from 1-year-old black walnut 
seedlings was used. 

The 1980 test was done with Forest Service volunteers at Carbondale, 
Illinois, the headquarters for all the Forest Service research with black walnut. 
This group of 19 volunteers included people who had never been exposed to black 
walnut, and others who had been working with the species up to 10 years. Six 
were tested with green bark from current year's growth and 15 were tested with 
brown, 1-year-old-bark from older trees; two were tested with both. 

The test procedure in 1979 and 1980 was as follows: 

1. Cut a piece of bark about V2-inch square and tape into close contact with 
bare skin (inside the arm) using surgical tape or masking tape. 

2. Leave in place for 4 hours. (If the area becomes uncomfortable prior to the 
recommended 4 hours completion time, remove patch and record length of time 
patch is in place and condition under which it was removed.) The bark patch was 
left in place from 1 to 24 hours on the Hoosier volunteers in the 1979 test. 

3. Observe and record reaction at patch site immediately after patch 
removal and 24 hours after patch removal. 

Results 
Questionnaire 

The response to the call for information in the Walnut Council Newsletter 
was minimal. We received only seven replies. However, from those respondents 
who did answer our questionnaire, a pattern did emerge. All those who had 
experienced reaction similar to those of the foresters were working with live 
walnut trees. Four were pruning; one was hulling nuts; one was logging; and one 
was removing vines from the trees (the vine remover may not be appropriate 
because the species of the vine is suspect). 

All of the episodes of contact dermatitis took place between the months of 
July and October. All respondents commented that the weather was very 



'Mention of trade names does not constitute endorsement of the products by the USDA Forest Service. 



100 Indiana Academy of Science 

hot — about 90° F — and very humid, and stated that they were perspiring freely 
at the time of infection. Areas affected included hands, forearms, and in one case, 
the waistline where sawdust had settled. 

The reactions of the respondents were all similar. Within an hour of exposure 
each reaction began as a reddened area which developed blisters over the exposed 
area. One respondent reported tissue swelling. Another reported coughing when 
the nuts were being cracked. All but one respondent had had previous exposure 
to black walnut. 

Patch Tests 

The immediate reaction noticed by most volunteers was a burning and 
itching sensation under the patch material. When the patch was removed the area 
on some was pinkish and tender to touch. After 24 hours, blisters had formed on 
the test area. The blisters disappeared on all volunteers within seven days. 

1979 Patch Test 

All six volunteers experienced some immediate reaction; five people had a 
burning sensation and one had an itching sensation. Twenty-four hours later four 
of those with the initial burning sensation had blistering. One of those with the 
initial burning sensation and the one with the initial itching sensation experienced 
no further reaction. 

1980 Patch Test 

The initial reaction for the six tested with green bark was: none for one, 
pinkish skin for two, pink and slightly raised skin for one, tender skin for one, and 
blistered skin for one. Twenty-four hours after the patches were removed four of 
the above had or developed blisters; those that didn't were the one with no initial 
reaction and the one with pink and raised skin. 

Brown bark seemed less toxic. Initial reaction for the 15 tested with brown 
bark was: none for eight, burning and pink skin for two, burning and slightly 
raised for two, slightly raised for one, pink and raised for one, and stinging for 
one. Skin condition twenty-four hours after patch removal was as follows: no reac- 
tion for the eight with no initial reaction; of those that had had an initial reaction, 
three cleared up and four developed blisters. Both volunteers tested by both 
green and brown bark patches suffered reactions under the green bark patches 
but were unaffected by the brown bark patches. 

Discussion 

The substance in black walnut that probably causes the dermatitis is juglone, 
5-hydroxy-l, 4-naththoquinone (6). Juglone is found virtually in all parts of the liv- 
ing black walnut tree — stems, leaves, fruits, and roots (3). Brissemoret and 
Combes (3), as well as Daglish (4), confirm, however, that there is a definite 
seasonal variation in the amount and potency of the chemical in the trees. Accord- 
ing to Daglish (4) the highest concentrations are found in the winter buds, green 
twigs, male catkins, and the husks of very young fruits. The seasonal variation of 
juglone is substantiated by the work of Lee and Campbell (7) which shows that 
there is a definite increase of juglone in the hulls and leaves in the months of July 
to September. 

Juglone is a known allelopathic agent. Black walnut has been known since 
ancient times to inhibit growth of trees and plants growing close by (9). It has 
been shown that hydrojuglone, a nontoxic substance, is oxidized by the air or 



Botany 101 

some other oxidizing substance (8). This oxidized form penetrates into the soil and 
selectively inhibits growth of nearby plants (4,7,9). 

This same oxidation process which causes allelopathy also causes dermatitis 
of animals and man. In 1905 Brissemoret and Combes (3) applied a pommade of 
juglone, lanoline, and petroleum jelly on the skin of a rabbit. The results were 
"blackening of the tegument, formation of blisters, slight edema, thickening and 
hypertrophy of the epidermis." In 1931, Louis Schwartz (10), senior surgeon at 
U. S. Public Health Services, recorded several incidents of dermatitis in a cabinet- 
making factory where Brazilian walnut wood was being used. The reactions 
described by Dr. Schwartz were similar in description to those demonstrated by 
the rabbit-edema — blistering and hypertrophy of the epidermis. 

In 1954, Dr. John M. Siegel (11) in Archives of Dermatitis and Syphilology 
gives a case history of a 44-year-old white male with complaints of burning, 
itching, and "blistering eruption in the finger webs" after a day of picking black 
walnuts. 

In 1937, R. Barniske (2) related a case history of a 36-year-old female, who 
experienced blistering and erythema of her hands after working with walnut 
husks. Barniske thought this reaction was secondary to juglone. His testing was 
done with material from "several weeks old and already brown walnut husk." The 
test results were negative, but he "attributed this to the fact that the reponsible 
toxins in question (tannic acids and juglone), due to their chemical characteristics, 
already were present in the oxydized, i.e., no longer active, form in the stored 
walnut husks." 

Arndt (1) distinguishes between "irritant contact dermatitis" (which he 
divides into mild and strong) and "allergic contact dermatitis" as follows: 

"Primary irritant contact dermatitis is a nonallergic reaction of the skin 
caused by exposure to an irritating substance. There are two types of irritants: (1) 
mild, relative or marginal irritants, which require repeated and/or prolonged con- 
tact to produce inflammation (i.e., soaps and detergents), (2) strong or absolute 
irritants, which are such damaging substances that they will injure skin immedi- 
ately on contact (weeds and alkalis)." 

"Allergic contact dermatitis is a manifestation of delayed hypersensitivity 
and results from the exposure of sensitized individuals to contact allergins. . . .The 
incubation period after initial sensitization to an antigen is 5 to 21 days, while the 
reaction time after subsequent re-exposures is 12 to 48 hours." 

According to Arndt (1) mild irritants produce erythema, microvesiculations 
and oozing, whereas strong irritants cause blistering, erosion, and ulcerations. On 
the other hand a typical allergic reaction consists of grouped or linear tense 
vesicles and blisters. It can be accompanied by severe edema. 

Conclusions and Recommendations 

Our work shows that the contact dermatitis experienced by people exposed 
to the juglone of black walnut is probably an irritant rather than an allergic react- 
ion because: (1) the reactions occurred immediately, (2) the reactions were blister- 
ings and erosions, and (3) the reactions occurred in most people who were exposed 
to juglone. This affirms the work of Siegel (11) who stated, "The absence of der- 
matitis on other exposed parts of the body and hands which were also heavily 
stained with walnut juice. . .points toward a primary irritant effect of the walnut 
juice as the cause rather than allergic sensitivity." Barniske (2) supports the 
premise that the chemical juglone is the primary irritant. 



102 Indiana Academy of Science 

If conditions are favorable, most people exposed to juglone will develop con- 
tact dermatitis. People are more sensitive to juglone poisoning during hot 
weather, especially when it is hot and humid. The two Forest Service people were 
working with walnut during unusually warm or hot humid weather and so were all 
of the respondents to the Walnut Council Newsletter. Conditions for inflamma- 
tion seem even more favorable when the victim perspires. 

To prevent juglone dermatitis: 

1. Avoid the juice from walnut stems, nuts, leaves, and roots. 

2. Avoid hot days for pruning walnut trees or hulling the nuts. 

3. Wear protective clothing: 

a. Rubber gloves when gathering or hulling nuts. 

b. Long sleeve shirt, buttoned at the throat, when pruning walnut trees. 

4. Wash off any walnut chips, or juice from the walnut hull, as soon as possi- 
ble. 

Literature Cited 

1. Arndt, Kenneth A., M.D. 1974. Manual of dermatological therapeutics. 377 p. 
Little, Brown and Company, Boston. 

2. Barniske, R. 1957. Dermatitis bulbosa, ausgelost durch den Saft gruner 
Walnussfruchtschalen (Juglons regia). p. 838-840. Dermatologische Wochen- 
schrift 135(8):189-192. Translated by Knud Clausen. 

3. Brissemoret, M. M. and R. Combes. 1905. Sur le juglon. Comptes renders del' 
Academies des Sciences 141:838. Translated by Johanna Clausen. 

4. Daglish, C. 1950. The determination and occurrence of a hydrojuglone glu- 
coside in the walnut. Biochem. J. (London) 47(l):458-462. 

5. Funk, David T. and Robert T. Williams. 1979. Can Juglone be hazardous to 
your health? Walnut Counc. Newsl. 6(2). 

6. Jesaitis, R. G. and A. Krantz. 1972. Juglone: an organic chemistry-ecology in- 
teraction experiment. J. Chem. Educ. 49:4-437. 

7. Lee, K. C, and R. W. Campbell. 1965. Nature and occurrence of juglone in 
Juglans nigra L. Hortscience 4:297-298. 

8. Marking, Leif L. 1970. Juglone (5-hydroxy-l, 4-naththoquinone) as a fish toxi- 
cant. Trans. Amer. Fish. Soc. 99(3):510-514. 

9. Rietveld, W. J. 1979. Ecological implications of allelopathy in forestry, p. 
91-112. In Proc. John S. Wright Forestry Conf., Purdue Univ., W. Lafayette, 
Indiana, Feb. 22-23, 1979. 

10. Schwartz, Louis. 1931. Dermatitis venenata due to contact with Brazilian 
walnut wood. Pub. Health Rep. 46:1938-1942. 

11. Siegel, John M., M.D. 1954. Dermatitis due to black v/alnut juice. A.M.A. 
Archives of Dermatology and Syphilology 70(6):511-513. 

Acknowledgements 

The gratitude of the authors is extended to those seven members of the 
Walnut Council who answered the questionnaire cited in this paper and to the 
twenty-five brave people who volunteered for the patch testing, especially Ed 
Hartsell and Mace Clark. Last, but not least, thanks also go to Dave Funk for his 
encouragement and support in this undertaking. 



Woody Vegetation of the Cass County Outdoor 
Education and Wildlife Area 

Gary E. Dolph, Jon Bolke, and Jill Rodaer 
Indiana University at Kokomo, Kokomo, Indiana 46902 

Introduction 

The Cass County Outdoor Education and Wildlife Area (Sec. 17, T 27 N, R 2 
E, Logansport Indiana Quadrangle) is a new park currently being developed on 
property located to the east of the Cass County Home. The vegetation found in 
the park is of two types. Along the central stream and the eastern ridge crest, the 
vegetation is relatively mature and could serve as a nature study area. Off the 
ridge crest to the east and west of the stream are a series of old abandoned fields 
in various stages of succession. Only the woody vegetation surrounding the cen- 
tral stream will be discussed in this paper. The authors hope that the information 
included in this paper will aid school teachers and hikers who are using the nature 
study area. 

Materials and Methods 

Two line transects were run roughly parallel to the stream bed. The ridge 
transect ran along the ridge crest from the northwestern to the southwestern cor- 
ner of the park. The transect was discontinuous to avoid areas of tree fall. The 
lowland transect ran along the eastern stream bottom except in the southern part 
of the park where it crossed the stream and ran along its western border. Each 
transect was 500 m long. Both the tree and shrub layers were sampled along each 
transect. Importance values for the dominant tree species were determined using 
the point-centered quarter method (5, 6). The sampling points were located at 20 
m intervals. The individual closest to the sampling point in each quarter whose 
diameter at ground level exceeded 4 in. (10 cm) was sampled. The shrub layer was 
sampled using 10 m sq quadrats. The 10 quadrats analyzed on each transect were 
located at 20 m intervals. All individuals taller than 1 m and having a diameter at 
ground level of less than 4 in. (10 cm) in the quadrat were recorded. The impor- 
tance value for each shrub species encountered was calculated using the method 
of Curtis and Mcintosh (1, 2). An analysis for stand disturbance was run also along 
the lowland transect (7). 

Results 

Stand disturbance analysis seemed to indicate that the vegetation of the 
stream bottom was not disturbed. Correlation analysis (8) indicated that the 
observed variation was not significantly different from a straight line (r = -0.9219). 
Other evidence at the site contradicted this conclusion, however. A logging road 
entered the stream valley at the southern end of the park. Numerous drag lines 
were encountered running up the ridges out of the stream valley. The majority of 
large diameter trees, particularly those on the ridge crest, were highly branched 
and not suitable for logging. These trees had branching patterns characteristic of 
trees grown in the open. Finally, a large number of escaped ornamentals, 
including Lonicera tatarica, Berberis thunbergii, and Malus, were found in the 
stream valley. The area was logged a considerable time ago, and the stream valley 
vegetation has apparently achieved a normal size distribution in all but the 
largest size classes. 

103 



104 Indiana Academy of Science 

A total of 37 different species were encountered along the lowland and ridge 
transects. In addition to these species, eight others were encountered while walk- 
ing through the woods. The additional species encountered were Rhus radicans, 
Ribes cynosbati, Viburnum dentatum, Parthenocissus quinque folia, Rubus flag- 
gellaris, Populus deltoides, Fagus grandifolia, and Lonicera japonico. 

Except for the dominance of Ulmus rubra along both the lowland and ridge 
transects (Tables 1 and 2), the canopy layers in the stream valley and on the ridge 
crest were quite distinct. Ulmus rubra and Juglans nigra were distributed 
throughout the length of the lowland transect. The other species were distributed 
somewhat more randomly. Asimina triloba and Plantanus occidentalis were con- 
fined to the northern end, while Prunus serotina, Celtis occidentalis, Fraxinus 
americana and Morus rubra were confined to the southern end of the lowland 
transect. Acer saccharum was found in the central area of the lowland transect. 
Malus was represented by a single individual. The distribution of canopy trees 
was irregular along the ridge transect and no one species was dominant along the 
entire length of the transect. Acer saccharum, Ulmus rubra, and Prunus serotina 
were most common on the southern part of the ridge; Sassafras albidum occurred 
exclusively in the central part; and Quercus rubra, Juglans nigra, and Ulmus 
rubra were the most abundant species at the northern end. The vegetation at the 
northern end of the ridge transect appeared to be the least disturbed of any 
vegetation in the park. 

The shrub layer varied considerably between the stream valley and the ridge 
crest (Tables 1 and 2). The shrub layer was most diverse in the stream valley. 
Thirty different species were encountered in the stream valley, while only 14 
were encountered along the ridge crest. All the additional species encountered 
while walking through the park occurred most frequently along the stream bot- 
tom. At the northern end of the lowland transect, the most common species were 
Prunus serotina, Asimina triloba, Berberis thunbergii, and Rosa Carolina Zan- 



Table 1. Importance values of the woody plants found along the ridge transect 

in the Cass County Outdoor Education and Wildlife Area. The species were 

identified using Deam (3, k). 

Species Canopy Layer Shrub Layer 

Ulmus rubra 

Acer saccharum 

Prunus serotina 

Quercus rubra 

Tilia americana 

Crataegus sp. 

Platanus occidentalis 

Juglans nigra 

Morus rubra 

Juglans cinerea 

Celtis occidentalis 

Sassafras albidum 

Quercus muhlenbergii 

Viburnum lentago 

Asimina triloba 

Lindera benzoin 

Rosa Carolina 4.6 

Carpinus caroliniana 2.5 

Viburnum rafinesquianum 2.5 



92.7 


27.4 


55.1 


156.3 


45.6 


59.4 


37.4 


6.9 


14.5 




12.0 


4.8 


11.4 




6.6 




6.5 


2.6 


5.7 




4.6 


7.1 


4.2 


7.3 


3.8 






7.9 




6.2 




4.6 



Botany 



105 



Table 2. Importance values of the woody plants found along the lowland 

transect in the Cass County Outdoor Education and Wildlife Area. The species 

were identified using Deam (3, k). 



Species 



Canopy Layer 



Shrub Layer 



Ulmus rubra 
Platanus occidentalis 
Juglans nigra 
Prunus serotina 
Celtis occidentalis 
Acer saccharum 
Crataegus sp. 
Asimina triloba 
Morus rubra 
Fraxinus americana 
Malus sp. 
Rosa Carolina 
Lindera benzoin 
Berberis thunbergii 
Fraxinus americana 
Sambucus canadensis 
Rubus allegheniensis 
Carya oralis 
Viburnum lentago 
Zanthoxylum americanum 
Lonicera tatarica 
Carya cordiformis 
Carpinus caroliniana 
Gleditsia triachanthos 
Corn us florida 
Ostrya virginiana 
Viburnum rafinesquianum 
Smilax rotundifolia 
Quercus rubra 
Aesculus glabra 
Vitis riparia 
Salix nigra 



132.5 


83.6 


32.5 


1.5 


22.0 


1.3 


22.0 


38.6 


20.6 


8.8 


16.8 


45.3 


16.6 


10.1 


11.4 


29.5 


10.8 


1.2 


10.7 




4.0 






18.0 




14.6 




9.9 




7.1 




3.1 




3.1 




2.8 




2.6 




2.6 




2.3 




1.7 




1.4 




1.3 




1.2 




1.2 




1.2 




1.2 




1.2 




1.2 




1.2 




1.2 



thoxylum americanum and Rubus allegheniensis were most common in the cen- 
tral area. Acer saccharum and Lonicera tatarica were most common at the 
southern end of the lowland transect. Unfortunately, Rhus radicans was very 
abundant throughout the stream valley. In addition, the percentage of plants in 
the shrub layer of the stream valley having spines or thorns was quite high, mak- 
ing any deviations from the regular nature trails very difficult. The low diversity 
of species in the shrub layer along the ridge crest parallels the low diversity that 
was found in the canopy layer. Acer saccharum, Ulmus rubra, and Prunus 
serotina were most common in the southern part of the transect. Sassafras 
albidum, due to the production of sucker shoots, was very common in the central 
portion. The remainder of the species occurred most frequently at the northern 
end of the ridge transect. The diversity on the ridge crest increases at the nor- 
thern end of the park and approaches that of the stream valley. 

Conclusions 

The lowland stream valley and adjacent ridge crests are well suited to serve 
as a nature study area. Most of the common species are visible from the nature 
trails. However, many people may be discouraged from wandering through the 
area due to the spiny nature of most of the shrub layer plants and the abundance 



106 Indiana Academy of Science 

of Rhus radicans. The vegetation outside of the study area is less suitable for 
nature study. Some of the old field area should be retained as an aid in studying 
plant succession, but the vast majority of this area could be modified for other 
purposes such as picnicking and athletics. 



Literature Cited 

1 . Curtis, J. T. and R. P. McIntosh. 1950. The interrelation of certain analytic and syn- 
thetic phytosociological characters. Ecology 31: 434-455. 

2. . 1951. An upland forest continuum in the prairie-forest border region of 

Wisconsin. Ecology 32: 476-496. 

3. Deam, C. C. 1932. Shrubs of Indiana. Dept. of Cons., Indianapolis. 380 pp. 

4. . 1953. Trees of Indiana. Dept. of Cons., Indianapolis. 330 pp. 

5. Dolph, G. E. 1976. A computer program for calculating the importance value of tree 
species in a sample stand. In: A. Schaeffer and L. Motsinger (eds.), Proceedings of the 
3rd Annual Conference on Instructional Computing Applications, pp. 39-53, Indiana 
University at South Bend, South Bend, Indiana. 231 pp. 

6. Mueller-Dombois, D. and H. Ellenberg. 1974. Aims and methods of vegetation 
ecology. John Wiley and Sons, Inc., New York. 547 pp. 

7. Schmelz, D. V. and A. A. Lindsey. 1965. Size-class structure of old-growth forests 
in Indiana. For. Sci. 11(3): 258-264. 

8. Sokal, R. R. and F. J. Rohlf. 1969. Biometry. W.H. Freeman and Company, San Fran- 
cisco. 776 pp. 



A Compilation of Plant Diseases and Disorders in Indiana— 1980 

Gail E. Evans-Ruhl, Donald H. Scott, Paul C. Pecknold 

Department of Botany and Plant Pathology 
Purdue University, West Lafayette, Indiana 47907 

Introduction 

This paper is a compilation of those plant diseases and disorders which were 
diagnosed at the Purdue Plant Diagnostic Clinic from January 1 through October 
30, 1980. Its purpose is to show which diseases and disorders are present in Indi- 
ana, their frequency, and whether they are increasing or decreasing. Ultimately, 
comparisons of yearly disease-disorder problems will enable us to (1) recognize the 
problems and initiate timely control efforts, (2) provide information for 
recommending plant varieties best suited for Indiana, and (3) give additional in- 
sight to plant diseases and disorders in the past, present, and future. 

Methods 

Plant specimens were submitted to the Plant Diagnostic Clinic from county 
extension agents, homeowners, growers, nursery operators and others. 
Specimens were diagnosed visually or by culturing the pathogen on selected 
media. Once diagnosed, appropriate control measures were given for each sample 
submitted. A breakdown of the total number of specimens handled from January 
1 through October 30, 1980 is given in Table 1. 

Table 1. Total Number of Specimens Handled from January 1 Through October 

30, 1980. 



Plant Species 

AGRONOMIC (21%> 
Corn 
Soybeans 
Small Grain 

Forage Grasses and Legumes 
Tobacco 

ORNAMENTAL (35%) 

Trees-Shade & Ornamental 
Shrubs and Groundcover 
Flowers 
House plants 

FRUIT (10% I 
Tree Fruit 
Small Fruit 

VEGETABLE (11%) 

TURFGRASS (5%) 

PLANT IDENTIFICATION (12%) 205 

FORWARDED TO ENTOMOLOGY (6%l 93 

Total 1650 688 555 109 210 

'Problem caused by an infectious disease causing agent eg fungus, bacterium, virus, mycoplasma, nematode. 
'Problem caused by noninfectious environmental stress e.g., wind, drought, heat, soil compaction. 
'Problem caused by herbicide or fertilizer misuse. 

107 











Number of 










Different 


Number of 








Infectious 


Samples 


Disease 1 


Disorder 2 


Chemical 3 Cai 


isal Agents 


161 


96 


45 


20 


16 


114 


60 


28 


26 


11 


57 


50 


7 





15 


20 


15 


5 





7 


2 


1 





1 


1 


362 


106 


238 


18 


32 


126 


72 


52 


2 


23 


43 


27 


9 


7 


14 


27 


9 


17 


1 


7 


103 


47 


48 


8 


17 


59 


32 


24 


3 


15 


189 


104 


63 


22 


40 


89 


69 


19 


1 


12 



108 



Indiana Academy of Science 



Results 

Weather related problems were commonplace in Indiana during the 1980 
growing season (Table 1). Wet weather occurred in the spring and was followed 
by below normal precipitation during the summer in the western half of the state. 
Precipitation approached normal in the eastern part. Above normal temperatures 
prevailed throughout the state in July, August, and September. 

Disease losses were relatively light during 1980, even though some infectious 
diseases became widespread in drought and heat-stressed plants. 

Tables 2-8 show the host plant, the disease or disorder diagnosed, the 
pathogen or cause of disorder, and the number of samples received. 

Table 2. Shade and Ornamental Trees— Diseases and Disorders 

Host Plant Number of 
Diseases and/or Disorders Causal Agent Samples 

Abies (FIR) 

Miscellaneous Disorders 
Poor Vigor 
Needle Tip Burn 

Acer (MAPLE) 
Anthracnose 
Wilt 
Canker 
Wetwood 

Miscellaneous Disorders 
Scorch 
Dieback 
Bark Shedding 
Herbicide Injury 
Frost Crack 
Chlorosis 

Acer (BOX ELDER) 

Miscellaneous Disorders 
Dieback 
Scorch 

Carya (HICKORY) 
Anthracnose 

Miscellaneous Disorder 
Scorch 

Catalpa (CATALPA) 

Miscellaneous Disorder 
Scorch 

Cercis (RED BUD) 
Wilt 

Miscellaneous Disorders 
Scorch 
Dieback 

Comus (DOGWOOD) 
Canker 
Leaf Spot 

Miscellaneous Disorders 
Scorch 
Dieback 
Herbicide Injury 



Stress factor(s) 

Heat, wind and drought 

Gloeosporium apocryptum 
Verticillium albo-atrum 
Unidentified 
Unidentified bacteria 

Heat, wind and drought 

Stress factor(s) 

Natural 

Spray drift 

Winter temperature extremes 

Nutrient deficiency 



Stress factor(s) 

Heat, wind and drought 

Gnomonia caryae 

Heat, wind and drought 

Heat, wind and drought 

Verticillium albo-atrum 

Heat, wind and drought 
Stress factor(s) 

Botryosphaeria sp. 
Unidentified fungus 

Heat, wind and drought 
Stress factor(s) 
Spray drift 



Botany 
Table 2- Continued 



109 



Crataegus (HAWTHORN) 

Cedar Hawthorn Rust 
Elaeagnus (RUSSIAN OLIVE) 

Canker 

Wilt 

Fraxinus (ASH) 
Anthracnose 
Canker 

Miscellaneous Disorders 
Scorch 
Dieback 
Chemical Injury 

Juglans (WALNUT) 

Miscellaneous Disorder 
Scorch 

Juniperus virginiana (RED CEDAR) 
Twig Blight 

Miscellaneous Disorders 

Dieback 
Liquidambar (SWEET GUM) 

Miscellaneous Disorders 
Scorch 
Dieback 

Liriodendron (TULIP TREE) 
Powdery Mildew 
Wilt 

Miscellaneous Disorders 
Scorch 

Leaf Yellowing/Spotting 
Chemical Injury 

Magnolia (MAGNOLIA) 

Miscellaneous Disorders 
Winter Damage 
Dieback 
Herbicide Injury 

Malus (CRABAPPLE) 
Scab 

Cedar-Apple Rust 
Fireblight 

Miscellaneous Disorders 
Scorch 
Dieback 

Picea (SPRUCE) 
Canker 
Needle Cast 

Miscellaneous Disorders 
Dieback 

Chemical Injury 
Needle Tip Burn 
Mechanical damage 

Pmus (PINE) 
Tip Blight 
Needle Cast 
Needle Blight 
Pinewood Nematode 



Gymnosporangium globosum 

Fusicoccum elaeagni 
Verticillium albo-atrum 

Gloeosporium aridum 
Unknown 

Heat, wind and drought 
Stress factor(s) 
Spray drift 



Heat, wind and drought 
Phomopsis juniperovora 
Stress factor(s) 



Heat, wind and drought 
Stress factor(s) 

Unidentified 
Verticillium albo-atrum 

Heat, wind and drought 

Natural 
Spray drift 



Desiccation 
Stress factor(s) 
Spray drift 

Venturia inaequalis 

Gymnosporangium juniperi-virginianae 
Erwinia amylovora 

Wind, heat and drought 
Stress factor(s) 

Cytospora kunzei 
Rhizosphaera kalkoffii 

Stress factor(s) 

Improper uses 

Heat, wind and drought 

Unidentified 

Diplodia pinea 
Lophodermium pinastri 
Dothistroma pirn 
Bursaphelenchus lignirolus 



110 



Indiana Academy of Science 
Table 2 — Continued 



Miscellaneous Disorders 
Decline 

Air Pollution Injury 
Winter Damage 
Needle Tip Burn 
Sooty Mold 
Yellowing/Autumn 
(previous yrs growth) 
Mechanical Damage 

Platanus (SYCAMORE) 
Anthracnose 



Poor drainage site-stress factor(s) 

Air pollutants 

Desiccation 

Heat, wind and drought 

Insect honeydew secretions 

Natural 

Unidentified 



Gnomonia veneta 
Populus (POPLAR, ASPEN, COTTONWOOD) 



Miscellaneous Disorders 
Dieback 

Prunus (PURPLE LEAF PLUM) 

Miscellaneous Disorder 
Winter Damage 

Prunus (FLOWERING ALMOND) 
Fireblight 

Prunus (ORIENTAL CHERRY) 

Miscellaneous Disorder 
Winter Damage 

Quercus (OAK) 
Anthracnose 
Oak Wilt 
Leaf Blister 
Leaf Spot 
Wetwood 

Miscellaneous Disorders 
Chlorosis (Pin Oak) 
Dieback 
Scorch 
Chemical Injury 

Robinia (LOCUST) 

Miscellaneous Disorders 
Scorch 
Dieback 

Salix (WILLOW) 
Canker 
Canker 
Canker 

Sorbus (MOUNTAIN ASH) 
Fireblight 
Leaf Spot 
Leaf Spot 

Miscellaneous Disorders 
Scorch 
Dieback 

Tilia (LINDEN) 

Miscellaneous Disorder 
Dieback 

Tsuga (HEMLOCK) 

Miscellaneous Disorder 
Sooty Mold 



Stress factor(s) 



Desiccation 



Erwinia amylovora 



Desiccation 

Gnomonia quercina 
Ceratocystis fagacearum 
Taphrina coerulescens 
Actinopelte dryina 
Unidentified bacteria 

Iron deficiency 
Stress factor(s) 
Wind, heat and drought 
Spray drift 



Wind, heat and drought 
Stress factor(s) 

Botryosphaeria ribis 
Cytospora chrysosperma 
Unidentified 

Erwinia amylovora 
Phyllosticta globigera 
Septoria sp. 

Heat, wind and drought 
Stress factor(s) 



Stress factor(s) 



Insect honeydew secretions 



Botany 
Table 2 — Continued 



111 



Ulmus (ELM) 

Dutch Elm Disease 
Black Spot 
Leaf Spot 

Miscellaneous Disorders 
Dieback 
Chlorosis 
Chemical Injury 



Ceratocystis ulmi 
Gnomonia ulmea 
Phyllosticta sp. 

Stress factor(s) 
Nutrient deficiency 
Spray drift 



Table 3. Ornamentals— Diseases and Disorders 



Hose Plant 
Disease and/or Disorder 



Causal Agent 



Number of 
Samples 



Ajuga (BUGLE-WEED) 

Miscellaneous Disorder 
Scorch 

Aloe (BURN PLANT) 

Miscellaneous Disorder 
Leaf Spot 

Amaryllis (AMARYLLIS) 
Red Blotch 

Amelanchier (SERVICEBERRYI 
Root Rot 

Aquilegia (COLUMBINE) 

Miscellaneous Disorder 

Leaf Spot/Malformation 

Begonia (BEGONIA) 
Dieback 

Berberis (BARBERRY) 

Miscellaneous Disorder 
Dieback 

Campanula (BELLFLOWER) 

Miscellaneous Disorder 
Bud Abortion 

Celosia (COCKSCOMB) 
Stem Rot 

Cissus (GRAPE IVY) 

Miscellaneous Disorder 
Chlorosis 

Clematis (CLEMATISl 

Miscellaneous Disorder 
Scorch 

Cotoneaster (COTONEASTER) 
Fireblight 

Chrysanthemum (CHRYSANTHEMUM! 
Stem Rot 
Stem Rot 
Stem Rot 
Root Rot 
Bacterial Blight 

Dianthus caryophyllus (CARNATION) 
Stem Rot 



Heat, wind and drought 

Improper cultural practices 
Stagonospora curtisii 
Armillaria sp. 

Environmental 
Botrytis sp. 

Improper site/poor drainage 

Excess water 
Rhizoctonia solani 

Improper cultural conditions 

Wind, heat and drought 

Erwinia amylovora 

Fusarium oxysporum 
Rhizoctonia solani 
Pythium sp. 
Pythium sp. 
Erwinia chrysanthemx 

Rhizoctoma solani 



112 



Indiana Academy of Science 
Table 3 — Continued 



Miscellaneous Disorder 
Stem Rot 

Dieffenbachia (DIEFFENBACHIA) 
Anthracnose Leaf Spot 

Miscellaneous Disorder 
Leaf Spot 

Euonymus (BURNING BUSH) 
Crown Gall 
Powdery Mildew 
Dieback/Canker 
Dieback 

Miscellaneous Disorder 
Dieback 
Herbicide Injury 

Euphorbia (SNOW ON THE MOUNTAINS) 
Root Rot 

Ficus (WEEPING FIG) 

Miscellaneous Disorder 
Leaf Spot 

Ficus (RUBBER PLANT) 
Anthracnose Leaf Spot 

Miscellaneous Disorder 
Edema 

Forsythia (GOLDEN BELLS) 
Crown Gall 

Miscellaneous Disorder 
Dieback 

Hedera (ENGLISH IVY) 

Miscellaneous Disorder 
Leaf Spot 

Hemerocallis (DAYLILLY) 
Dieback 

Miscellaneous Disorder 
Leaf Spot 

Hibiscus (ROSE-OF-SHARON) 
Root Rot 

Hosta (PLANTAIN-LILY) 
Leaf Spot 

Ilex (HOLLY) 
Leaf Spot 

Miscellaneous Disorders 
Winter Damage 
Chlorosis 

Juniperus (JUNIPER) 
Twig Blight 
Cedar-Apple Rust 
Cedar-Hawthorne Rust 

Miscellaneous Disorders 
Dieback 
Dieback 

Ligustrum (PRIVET) 

Miscellaneous Disorder 
Dieback 



High soluble salts 

Colletotrichum sp. 

Improper cultural conditions 

Agrobacterium tumefaciens 
Not identified 
Sclerotinia sclerotiorum 
Nectria/cold 

Stress factors 
Spray drift 

Rhizoctonia solani 

Improper cultural conditions 

Glomerella cingulata 

High soil moisture-retarded transpiration 

Agrobacterium tumefaciens 

Stress factors 

Improper cultural care 

Botrytis sp. 

Improper cultural conditions 

Not identified 

Phyllosticta sp. 

Phyllosticta spp. 

Desiccation 
Iron deficiency 

Phomopsis juniperovora 

Gym.no sporangium juniperi-virginianae 

Gymno sporangium globosum 

Shading out 
Dog injury 



Heat, wind and drought 



Botany 
Table 3— Continued 



113 



Lonicera (HONEYSUCKLE) 

Miscellaneous Disorder 
Scorch 
Winter Damage 

Orchid (ORCHID) 

Miscellaneous Disorder 
Leaf Spot 

Pachysandra (PACHYSANDRAI 

Leaf Blight 
Root/Stem Canker 

Miscellaneous Disorder 
Dieback 

Paeonia (PEONY* 

Red Spot (measles) 
Botrytis Blight 
Crown Rot 
Leaf Spot 

Miscellaneous Disorder 
Herbicide Injury 

Parthenocissus (BOSTON IVY) 
Leaf Spot 

Pelargonium (GERANIUM) 
Stem Canker 
Blackleg 
Bacteria] Blight 

Miscellaneous Disorders 
Chlorosis 
Root Rot 

Persea (AVOCADO) 

Edema 
Petunia (PETUNIA) 

Miscellaneous Disorder 
Root Rot 

Philodendron (PHILODENDRONI 
Anthracnose 

Miscellaneous Disorder 
Leaf Spot 

Polypodium (FERN) 
Root/Stem Canker 

Miscellaneous Disorder 
Root Rot 
Leaf Spot 

Pyracantha (FIRETHORN) 
Scab 



Heat, wind and drought 
Desiccation 



Improper cultural conditions 

Volutella pachysandrae 
Rhizoctonia solani 

Stress factor(s) 

Cladosporium paeoniae 
Botrytis cinerea 
Phytophthora cactorum 
Alternaria sp. 

Spray drift 

Guignardia bidwelli 

Rhizoctonia solani 
Pythium sp. 
Xanthomonas pelargonii 

Improper cultural conditions 
Soluble salt injury 

High soil moisture-retarded transpiration 



Soluble salt injury 

Co lie to trie hum sp. 

Improper cultural conditions 

Rhizoctonia solani 

Soluble salt injury 
Spore formation 



Fusicladium pyracanthae 
Rhododendron (AZALEA and RHODODENDRON) 



Crown Rot 
Leaf Spot 
Leaf Spot 

Miscellaneous Disorders 
Winter Scorch 
Chlorosis 

Rosa (ROSE) 
Blackspot 



Phytophthora spp. 
Pestalotia sp. 
Coryneum sp. 

Desiccation 

Improper cultural conditions 

Diplocarpon rosae 



114 



Indiana Academy of Science 
Table 3 — Continued 



Brand Canker 
Blossom Blight 

Miscellaneous Disorders 
Herbicide Injury 
Cold Injury 



Coniothyrium wemsdorffiae 
Botrytis cinerea 

Spray drift 

Low temperatures 



Saintpaulia (AFRICAN VIOLET) 

Miscellaneous Disorders 
Root Rot 
Leaf Spot 

Salix (PUSSY WILLOW) 
Twig Blight 

Sckefflera (AUSTRALIAN UMBRELLA 

Miscellaneous Disorders 
Leaf Spot 
Root Rot 

Sinningia (GLOXINIA) 
Corm Rot 

Syringa (LILAC) 

Powdery Mildew 
Bacterial Blight 

Miscellaneous Disorders 
Scorch 
Herbicide Injury 

Tagetes (MARIGOLD) 

Miscellaneous Disorder 

Low Germination Rate 

Taxus (YEW) 

Root Rot-Dieback 

Miscellaneous Disorders 
Herbicide Injury 
Winter Damage 
Dieback 

Thuja (ARBORVITAE) 
Twig Dieback 
Root Rot 

Miscellaneous Disorders 
Winter Injury 
Inner Leaf Browning/Autumn 

Tradescantia (WANDERING JEW) 

Miscellaneous Disorder 
Leaf Spot 

Tulipa (TULIP) 

Miscellaneous Disorder 
Chemical Injury 

Viburnum (VIBURNUM) 

Miscellaneous Disorder 
Scorch 

Vinca (PERIWINKLE) 
Stem Blight 

Yucca (YUCCA) 
Leaf Spot 



Soluble salt injury 
Improper cultural conditions 

Phomopsis sp. 
TREE) 

Improper cultural conditions 
Overwatering 

Rhizoctonia solani 

Microsphaera alni 
Pseudomonas syringae 

Heat, wind and drought 
Spray drift 



Poor seed quality 
Phytophthora-poor drainage complex 

Spray drift 
Desiccation 
Dog injury 

Phomopsis juniperovora 
Phytophthora sp. 

Desiccation 

Natural phenomenon 

Improper cultural conditions 

Improper use 

Wind, heat and drought 
Phoma exigua var.exigua 
Coniothyrium concentricum 



22 



Botany 
Table 3 — Continued 



115 



Zinnia (ZINNIA) 

Powdery Mildew 
Alternaria Blight 



Erysiphe cichoracearum 
Alternaria zinniae 



Table 4. Fruit Trees— Diseases and Disorders 



Host Plant 
Disease and/or Disorder 



Causal Agent 



Number of 
Samples 



Malus sylvestris (APPLE) 
Crown Rot 
Scab 

Cedar-Apple Rust 
Quince Rust 
Fire Blight 
Frogeye Leaf Spot 
Canker 
Sooty Blotch 
Flyspeck 
Blister Spot 
Black Rot 

Miscellaneous Disorders 
Fruit Crack 
Dieback 
Measles 

Chemical Injury 
Frost Crack 
Scorch 
Bitter Pit 
Moldy Core 

Prunus americana (PLUM) 
Black Knot 

Miscellaneous Disorders 
Cold Injury 
Chemical Injury 

Prunus armeniaca (APRICOT) 
Bacterial Leaf Spot 
Scab 
Brown Rot 

Miscellaneous Disorders 
Chemical Injury 

Prunis avium (CHERRY) 
Cherry Leaf Spot 
Brown Rot 

Miscellaneous Disorders 
Scorch 
Root Rot 
Dieback 
Fruit Rot 

Prunis persica (PEACH) 
Bacterial Spot 
Scab 

Miscellaneous Disorders 
Scorch 
Root Rot 



Phytophthora cactorum 

Venturia inaequalis 

Gymnosporangium juniperi-virgiaanae 

Gymnosporangium clavipes 

Erwinia amylovora 

Physalospora obtusa 

Phoma sp. 

Gloeodes pomigena 

Microthyriella rubi 

Pseudomonas syringae 

Physalospora obtusa 

Water fluctuation extremes 
Stress Factor(s) 
Manganese toxicity 
Improper use 

Winter temperature extremes 
Heat, wind and drought 
Unknown — physiological 
Improper calyx closing 

Dibotryon morbosum 

Low temperatures 
Improper use 

Xanthomonas pruni 
Cladosporium carpophilum 
Monilinia fructicola 

Improper use 

Coccomyces hiemalis 
Monilinia fructicola 

Wind, heat and drought 
Wet site location 
Stress factor(s) 
Improper pollination 

Xanthomonas pruni 
Cladosporium carpophilum 

Wind, heat and drought 
Wet site location 



116 



Indiana Academy of Science 
Table 4 — Continued 



Prunus persica var. nectarina (NECTARINE) 
Bacterial Leaf Spot 
Scab 



Pyrus communis (PEAR) 
Fireblight 
Leaf Blight 

Miscellaneous Disorders 
Scorch 

Chemical Injury 
Poor Growth 



Xanthomonas pruni 
Cladosporium carpophilum 



Erwinia amylovora 
Fabraea maculata 



Heat, wind and drought 
Improper use 
Improper fertility 



Table 5. Small Frutis— Diseases and Disorders 



Host Plant 
Diseases and/or Disorder 



Causal Agent 



Number of 
Samples 



Fragaria grandiflora (STRAWBERRY) 
Black Root Rot 

Leaf Scorch 
Leaf Blight 
Gray Mold Rot 
Slime Mold 

Miscellaneous Disorders 
Scorch 
Root Rot 

Ribes grossularia (GOOSEBERRY) 
Powdery Mildew 

Ribes sativum (CURRANT) 
Leaf Mottle 

Rubus (RASPBERRY) 
Anthracnose 
Crumbly Berry 
Verticillium Wilt 
Leaf Spot 
Rust 
Leaf Curl 

Miscellaneous Disorders 
Cold Injury 

Vaccinium (BLUEBERRY) 

Miscellaneous Disorders 
Cold Injury 
Scorch 

Vitis (GRAPE) 
Black Rot 
Leaf Curl 
Downy Mildew 
Powdery Mildew 

Miscellaneous Disorders 

Herbicide Injury 
Cold Injury 
Improper Ripening 



Specific pathogen(s) 
not known 

Diplocarpon earliana 
Dendrophoma obscurans 
Botrytis cinerea 
Physarum cinereum 

Heat, wind and drought 
Insect-borer 

Sphaerotheca mors-uvae 

Virus 

Elsinoe veneta 

Virus 

Verticillium albo-atrum 

Septoria darrowi 

Gymnoconia peckiana 

Virus 

Late freezes 



Low temperatures 
Site location 

Guignardia bidwelli 
Virus 

Plasmopara viticola 
Uncinula necator 



Spray drift 

Low temperatures 

Lack of CHO accumulation 





Botany 


117 


Table 6. 


Turf— Diseases and Disorders 




Host Plant 




Number of 


Diseases and/or Disorder 


Causal Agent 


samples 


Poa pratensis (BLUEGRASS) 






Leaf Spot 


Helminthosporium spp. 


32 


Fusarium Blight 


Fusarium roseum complex 


4 


Dollar Spot 


Sclerotinia komoeocarpa 


2 


Stripe Smut 


Ustilago striiformis 


1 


Flag Smut 


Urocystis agropyri 


1 


Rhizoctonia (warm weather) 


Rhizoctonia solani 


5 


Slime Mold 


Physarum cinereum 


5 


Cottony Blight 


Pythium aphanidermatum 


5 


Anthracnose 


Colletotrichum graminicola 


3 


Stunt Nematode 


Tylenchorhynchus sp. 


1 


Lesion Nematode 


Pratylenchus sp. 


1 


Fairy Ring 


Basidiomycetes 


2 


Miscellaneous Disorders 






Chemical Injury 


Improper use 


1 


Excessive Thatch 


Improper cultural care 


8 


Scorch 


Drought 


7 


Agrostis tenuis (BENTGRASS) 






Snowmold 


Typhula itoana 


1 


Rhizoctonia (Cool weather) 


Rhizoctonia sp. 


3 


Blue wilt/dieback 


unknown 


7 



Table 7. Vegetables— Diseases and Disorders 



Host Plant 
Diseases and/or Di 



>rder 



Causal Agent 



Number of 
Samples 



Allium cepa (ONION) 
Damping Off 

Asparagus officinalis (ASPARAGUS) 
Needle Blight 
Rust 

Beta vulgaris (BEET) 
Crater Rot 

Brassica oleracea var. botrytis (CAULIFLOWER) 



Pythium sp. 

Cercospora asparagi 
Puccinia asparagi 

Rhizoctonia solani 



Downy Mildew 
Wirestem 

Brassica oleracea var. capitata (CABBAGE) 
Black Rot 

Miscellaneous Disorders 
Leaf Spot 
Chemical Injury 

Brassica rapa (TURNIP) 

Miscellaneous Disorder 
Root Girdling 

Brassica ruvo (BROCCOLI) 
Clubroot 

Capsicum frutescens (PEPPER) 
Bacterial Spot 
Damping Off 
Wilt 
Leaf Mottle 



Peronospora parasitica 
Rhizoctonia solani 



Xanthomonas campestris 



Physiological 
Spray drift 



Soluble salts 



Plasmodiophora brassicae 

Xanthomonas vesicatoria 
Pythium sp. 
Fusarium sp. 
Unidentified virus 



118 



Indiana Academy of Science 
Table 7 — Continued 



Miscellaneous Disorders 
Leaf Damage 
Leaf Yellows and Curls 
Fruit Injury 
Chemical Injury 

Citrullus vulgaris (WATERMELON) 
Fusarium Wilt 

Miscellaneous Disorders 
Chemical Injury 
Leaf Damage 
Chlorosis 

Cucumis melo (CANTALOUPE) 
Downy Midlew 
Leaf Spot 
Bacterial Wilt 

Miscellaneous Disorders 
Leaf Damage 
Chemical Injury 

Cucumis sativus (CUCUMBER) 
Mosaic 
Anthracnose 
Wilt 
Slime Mold 

Miscellaneous Disorders 
Leaf Damage 

Cucurbita moschata (PUMPKIN) 
Wilt 

Miscellaneous Disorder 
Chemical Injury 

Cucurbita pepo (ZUCCHINI) 

Miscellaneous Disorder 
Blossom End Rot 

Ipomoea batatas (SWEET POTATO) 

Scurf 

Soil Rot/Pox 
Lactuca sativa var. crispa (LEAF LETTUCE) 

Bottom Rot 

Lycopersicon esculentum (TOMATO) 
Septoria Leaf Spot 
Early Blight 
Stem Canker 
Fusarium Wilt 



Cold injury 
Nutrient deficiency 
Sunscald 
Salt toxicity 

Fusarium oxysporum 

Spray drift 
Wind injury 
Nutrient deficiency 

Pseudoperonospora cubensis 
Alternaria cucumerina 
Erwinia tracheiphila 

Wind injury 
Spray drift 

Cucumber mosaic virus 
Colletotrichum lagenarium 
Pythium sp. 
Physarum cinereum 

Wind burn 

Fusarium sp. 

Spray drift 

Physiological 



Late Blight 

Black-Dot-Root-Rot (Hydroponics) 

Sclerotium Wilt 

Bacterial Spot 

Bacterial Speck 

Anthracnose (fruit) 

Mosaic 

Damping Off (Hydroponics) 

Miscellaneous Disorders 
Chemical Injury 
Chemical Injury (hydroponics) 
Blossom End Rot 
Walnut Wilt 



Monilochaetes infuscans 
Streptomyces ipomoea 

Rhizoctonia solani 

Septoria lycopersici 
Alternaria solani 
Rhizoctonia solani 
Fusarium oxysporium f. sp. 
lycopersici 

Phytophthora infestans 
Colletotrichum coccodes 
Sclerotinia sclerotiorum 
Xanthomonas vesicatoria 
Pseudomonas tomato 

Colletotrichum coccodes 
Unidentified virus 
Pythium aphanidermatum 



Spray drift 

Heater fumes 

Physiological 

Walnut tree excretions 



Botany 
Table 7 — Continued 



119 



Leaf Yellowing and Curling 
(Hydroponics) 
Leaf Necrosis 
Corky Root 
Fruit Injury 
Wilt 
Leaf Roll 

Pastinaca sativa (PARSNIP) 

Miscellaneous Disorder 

Minimal Root Development 

Phaseolus vulgaris (SNAP BEAN) 
Root Rot 
Root Rot 
Root Rot 
Anthracnose 
Stem Rot 
Common Blight 

Miscellaneous Disorders 
Chemical Injury 
Leaf Yellowing/Bronzing 
Stem Damage 
Leaf Damage 

Pisum sativum (PEA) 
Bacterial Blight 
Sooty spot 

Rheum spp. (RHUBARB) 
Ascochyta Leaf Spot 
Anthracnose Leaf Spot 
Crown Rot 

Solarium melongena (EGGPLANT) 
Wilt and Yellow 
Yellow Fruit Spot 

Solatium tuberosum (POTATO) 
Tuber Rot 
Bacterial Soft Rot 
Rhizoctonia 
Late Blight 
Common Scab 
Blackleg 

Miscellaneous Disorders 
Hollow Heart 
Enlarged lenticels 



Nutrient imbalance 

Wind injury 

High soluble salt level 

Sunscald 

Lightening 

Physiological 



Nutritional imbalance 



Rhizoctonia solani 

Fusarium sp. 

Pythium sp. 

Colletotrichum lindemuthianum 

Sclerotinia sclerotiorum 

Xanthomonas phaseoli 

Spray drift 

Mites 

Hail 

Wind injury, sunscald 

Pseudomonas pisi 
Heterosporium sp. 

Ascochyta rhei 
Colletotrichum erumpens 
Phytophthora sp. 

Cold damage 
Sunscald 



Fusarium sp. 
Erwinia carotovora 
Rhizoctonia solani 
Phytophthora infestans 
Streptomyces scabies 
Erwinia atroseptica 

Heat injury 
Excess water 



Table 8. Agronomic Crops— Diseases and Disorders 



Host Plant 
Diseases and/or Disorder 



Causal Agent 



Number of 
Samples 



Triticum (WHEAT) 

Root Rot (see below) 

Rhizoctonia 

Take-All 

Fusarium Root Rot 
Wheat Spindle Streak 
Barley Yellow Dwarf 



Various (see below) 
Rhizoctonia solani 
Ophiobolus graminis 
Fusarium spp. 

Wheat Sprindle Streak Virus 
Barley Yellow Dwarf Virus 



120 



Indiana Academy of Science 
Table 8 — Continued 



Septoria Glume Blotch 

Scab 

Septoria Leaf Blotch 

Helminthosporium Leaf Spot 

Powdery Mildew 

Rhizoctonia Sharp Eye 

Loose Smut 

Miscellaneous Disorders 

Improper Root Development 

Chlorosis 

Leaf Discoloration 

A vena (OAT) 
Halo Blight 
Blasting 

Hordeum (BARLEY) 
Loose Smut 

Zea (Dent Corn) 

Seedling Blights (see below) 
Gib Seedling Blight 
Pythium 

Anthracnose 

Northern Corn Leaf Blight 
Northern Corn Leaf Spot 
Common Smut 
Common Rust 
Stewart's Blight 

Ear Rots (see below) 
Fusarium Kernel Rot 
Fusarium Kernel Rot 
Kernel Rot 
Kernel Rot 

Stalk Rots (see below) 
Fusarium Stalk Rot 
Gib Stalk Rot 

Helminthosporium Stalk Rot 
Southern Corn Leaf Blight 
Holcus Spot 

Miscellaneous Disorders 
Tall corn/Short corn 
Chemical Injury 
Chlorosis/striping 
Root restriction 
Silvering of leaves 
Clubbed Roots 

Glycine (SOYBEAN) 

Rhizoctonia Root Rot 

Phytophthora Root Rot 

Pythium Root Rot 

Bacterial Blight 

Pod & Stem Blight 

Brown Stem Rot 

Purple Seed Stain 

Anthracnose 

Charcoal Rot 

Brown Spot 

Soybean Cyst Nematode 



Septoria nodorum 
Gibberella zeae 
Septoria tritici 
Helminthosporium sativum 
Erysiphe graminis 
Rhizoctonia solani 
Ustilago tritici 



Too shallow planting 
Nutrient Deficiency 
Frost 



Pseudomonas coronafaciens 
Fusarium sp. 



Ustilago nuda 



Various (see below) 

Gibberella zeae 

Pythium aphanidermatum 

Co lie to trie hum graminicola 
Exserohilum turcicum 
Bipolaris carbonum (race II) 
Ustilago maydis 
Puccinia sorghi 
Erwinia stewartii 

Various (see below) 
Fusarium moniliforme 
Gibberella zeae 
Penicillium sp. 
Celphalosporium sp. 

Various (see below) 
Fusarium moniliforme 
Gibberella zeae 
Bipolaris carbonum 
Bipolaris maydis (Race 0) 
Pseudomonas syringae 



Environmental factors 
Wet Weather 
Nutrient deficiency 
Soil compaction 
Cold injury 
Too deep planting 

Rhizoctonia solani 

Phytophthora megasperma var. sojae 
Pythium aphanidermatum 
Pseudomonas glycinea 
Diaportke phaseolorum var. sojae 
Cephalosporium gregatum 
Cercospora kikuchii 
Colletotrichum graminicola 
Macrophomina phaseolina 
Septoria glycines 
Heterodera glycines 



Botany 
Table 8 — Continued 



121 



Miscellaneous Diseases and Disorders 
Chemical Injury 
Chlorosis 
Root Restriction 
Leaf Discoloration 
Various Problems 
Leaf Bronzing/Defoliation 

Trifolium (CLOVER) 

Root Rot 
Miscellaneous Disorder 

Leaf Discoloration 
Medicago (ALFALFA) 

Sclerotinia Crown & Stem Rot 

Downy Mildew 

Spring Blackstem 

Leptosphaerilina Leaf Spot 

Rhizoctonia Root Rot 

Anthracnose 

Miscellaneous Disorders 
Leaf Discoloration 
White edges on leaves/ 
curling 

Nicotiana (TOBACCO) 
Blue Mold 

Miscellaneous Disorder 
Chemical Injury 



Various causes 
Nutrient deficiency 
Soil compaction 
Sunscald 

Environmental factors 
Mites 

Fusarium sp. 

Environmental factors 

Sclerotinia trifoliorum 
Peronospora trifoliorum 
Phoma medicaginis 
Leptosphaerulina briosiana 
Rhizoctonia solani 
Colletotrichum trifolii 

Environmental factors 
Frost 



Peronospora tabacina 
Improper use 



Shade and Ornamental Trees 

Diseases: As in the past years (2, 3) anthracnose of sycamore, maple, ash and 
white oak was the most common leaf disease of shade trees (Table 2). The late 
spring infection of sycamore resulted in excessive leaf drop during May and early 
June. Actinopelte leaf spot of oak was prevalent during late summer in those 
areas of the state which experienced excessive rainfall during August. Ver- 
ticillium was isolated from a number of shade trees, e.g. maple, catalpa, and ash, 
but was most frequently found on hard maple. Diplodia tip blight was common 
throughout the state, causing lower limb death in Scotch, Austrian and Red pine. 
A disease of pine trees, pinewood nematode, caused by the nematode, Bur- 
saphelenchus lignicolus, was recorded for the first time in Indiana. The disease 
was found in 8 counties of Indiana. It appears this disease has been present in 
Indiana, and throughout the Midwest, for a number of years but was not detected. 

Disorders: Maple trees throughout Indiana had an exceptionally heavy seed set 
which resulted in very sparse growth during the early spring months. The heavy 
seed set was attributed to a combination of the previous mild winter and lack of 
late spring freezes. A lower branch dieback of ash trees was noted in southern 
Indiana. Cause of the dieback was associated with stem cankering from anthrac- 
nose plus severe drought conditions. Leaf scorch was prevalent from late May 
through the summer. Early spring scorch was especially severe due to dry, hot, 
windy periods during late May and early June. Along with scorch, tree decline 
continued to be the most frequent problem of shade trees, especially maples. Site 
location and weather stress appeared to be major factors contributing to decline. 



122 Indiana Academy of Science 

Ornamentals 

Diseases: Juniper tip blight occurred in epidemic proportion in many areas of the 
state (Table 3). Current season growth was especially hard hit, however, older 
(mature) growth was relatively unaffected. Powdery mildew was prevalent on a 
number of ornamentals (lilac, rose, euonymus) during the late summer-early fall 
period. As in past years (2, 3), numerous samples of yew (Taxus) were received 
showing general dieback symptoms (Table 3). Yew dieback was most commonly 
associated with root injury due to wet site locations and/or Phytophthora sp. 

Disorders: The majority of disorders were related to a combination of poor site 
location along with drought stress. 

Tree Fruits 

Diseases: Apple scab, rust and fire blight were the most common diseases on 
apple (Table 4). Fire blight was most severe in the LaPorte fruit-growing area. 
Also on apple, collar rot, caused by Phytophthora cactorum, was more damaging 
than in previous years. Bacterial spot of peach and nectarine caused more damage 
than in recent years, however, damage was generally confined to leaves with little 
fruit infection. 

Disorders: Premature fruit drop of cherry, apricot, and plum was a major 
disorder. Cause of the premature drop was related to several factors, e.g. heavy 
fruit set, poor pollination, and insect injury. Many stone fruits showed decline 
symptoms as a result of the cold injury during previous winters (1977 thru 1979). 

Small Fruits 

Diseases: Bramble diseases were especially prevalent during 1980 (Table 5). 
Anthracnose and orange rust were common. Orange rust on blackberries has been 
especially widespread both this year and last year. Verticillium wilt of rasp- 
berries was frequently diagnosed as a problem in areas which previously had been 
planted in solanaceous crops. Though black root rot was the most common root 
problem of strawberry, fewer samples were received than in previous years (2,3). 
Black rot and downy mildew were the two most common grape diseases but were 
infrequent. 

Disorders: Cold injury continued to be a major disorder of brambles. While the 
past winter was mild, cold injury from the previous two winters continued to 
cause plant dieback; however, dieback was not as excessive as during the 1979 
growing season. 

Uneven ripening of grapes was reported frequently in the early part of the 
fall season. This disorder was attributed to a lack of carbohydrate accumulation 
which was apparently due to various factors such as excessive fruit crops (too 
little foliage), lack of adequate light intensity, high humidity, overpruning, 
powdery and downy mildew and an overall lack of functional foliage. 

Turfgrasses 

Diseases: Helminthosporium leaf spot and Helminthosporium melting out caused 
by Helminthosporium spp. were the most widespread and serious diseases of Ken- 
tucky bluegrass (Table 6). Helminthosporium melting out coupled with excessive 
thatch and environmental stress were responsible for killing large areas of turf in 
many lawns. Sporocarps of fairy ring fungi were common. An unidentified prob- 
lem occurred on at least three Indiana golf courses on Toronto C-15 bentgrass. 
The problem first appeared as a blue-wilt of lower leaf tips and progressed to 



Botany 123 

entire leaves and then all leaves of an individual plant. Affected leaves then 
yellowed and died. Affected plants occurred in small circular patches (1 to 3 inches 
in diameter), similar to dollar spot. Individual patches often coalesced killing 
large, irregular areas of turf. Laboratory studies failed to confirm the causal 
agent. Cool weather brown patch caused by Rhizoctonia was observed on both 
bentgrass and bluegrass in early spring. 

Disorders: Hot, dry weather resulted in poor root development and often caused 
plant death, especially where plants were further stressed by excessive thatch or 
disease. 

Vegetables 

Diseases: Specimens of cucurbit crops, snap bean, tomato and potato comprised 
the bulk of vegetable crops received (Table 7). Downy mildew was prevalent on 
muskmelon, as in past years (2,3). However, other foliar leaf spots were not signifi- 
cant. Heavy rainfall early in the growing season followed by widespread drought- 
like conditions, contributed to a high incidence of "first crop" blossom-end rot of 
tomato, pepper and squash. Bacterial spot (Xanthomonas vesicatoria) was a prob- 
lem early in the season on both pepper and tomato crops due to heavy rainfall and 
ample inoculum dispersal. An increased incidence of Fusarium and Rhizoctonia 
root rot of garden snap beans and sweet potato scurf (Monilochaetes infuscans) in- 
dicates the need for garden rotation and the use of resistant varieties where 
available (1). Hydroponic tomato operations in Indiana still are plagued with 
Pythium and black dot root rot. 

Disorders: As in previous years (2,3) a high number of vegetable samples were 
diagnosed as chemical injury. The majority of specimens originated in home 
gardens exposed to herbicide drift from a variety of sources. Clippings of 
herbicide-treated grass, used as a mulch around vegetable plants, also caused 
many injury problems. Environmental factors such as sun scald and wind injury 
caused a large share of scorch symptoms found on vegetable foliage and fruit. 

Agronomic Crops 

Agronomic Diseases: Damaging weather conditions were directly or indirectly 
related to an increased severity of several agronomic crop diseases (Table 8). 

Diseases— Wheat: Wheat spindle streak was widespread throughout the state in 
early spring, but yield losses were estimated to be less than 3 bu/acre. 

Septoria leaf blotch and powdery mildew were widespread with early spring rain, 
but they failed to develop to damaging proportions as drier weather conditions 
prevailed in late spring. Scab was widespread throughout the state, with some 
southeastern Indiana fields reaching up to 50% infection. Minor outbreaks of 
loose smut and, to a greater extent, bunt were reported. 

Disorders: The most common disorder was chlorosis due to nitrogen deficiency. 
Nitrogen loss was especially acute in southwestern Indiana. 

Disease — Oats: Halo blight caused by Pseudomonas coronafaciens was 
widespread. It was more prevalent than it has been in many years, but yield 
losses were minimal. 

Diseases — Corn: Seedling blights were common in early planted corn or where 
plants were stressed by soil compaction. Early season anthracnose leaf blight was 
widespread throughout the state in fields where corn followed corn. This phase of 
the disease was observed to be more severe than any previous year at the 3 to 6 
leaf stage of plant development. This stage of the disease did not affect yields, 



124 Indiana Academy of Science 

even though some anthracnose leaf blight was noted after tasseling. Holcus spot 
was observed in northern Indiana, but the disease did not develop to damaging 
proportions. Southern corn leaf blight, anthracnose leaf blight, Stewart's 
disease — leaf blight phase, and common rust were observed in numerous fields, 
but disease development was light. Minor yield losses from leaf blights occurred 
only in a few eastern and southwestern Indiana corn fields. Stalk rots were 
widespread and severe throughout the state. Some fields had up to 90% stalk rot. 
Ideal harvest conditions permitted early harvest, and yield losses from lodged 
corn was held to a minimum. Yield losses from stalk rots could have been severe 
with less than ideal harvest conditions. 

Disorders: The tall corn-short corn syndrome, caused by any factor that impedes 
root development, was a widespread and common complaint (Table 8). Soil com- 
paction appeared to be a major factor in the development of this problem. Con- 
siderable dinitroaniline carryover damage was observed, but this was felt to have 
been accentuated by the soil compaction problems. Drought and/or high 
temperatures caused some pollination problem. 

Disease— Soybeans: Phytophthora and Pythium seedling blights were severe and 
reduced stands in areas where excessive rainfall occurred shortly after planting. 
Rhizoctonia root rot was common but yield losses were slight. The soybean cyst 
nematode was found in additional fields in White and Cass counties. Bacterial 
blight and brown spot were common. Bacterial blight did not reduce yields. Fields 
severely affected with brown spot were prematurely defoliated and suffered some 
yield loss. Charcoal rot was commonly found in fields stressed by hot, dry 
weather. 

Literature Cited 

1. Chupp, Charles and A. F. Sherf. 1960. Vegetable Diseases and Their Con- 
trol. The Ronald Press Co., New York, 693 pp. 

2. Pecknold, P. C., W. R. Stevenson, and D. H. Scott. 1974. A Compilation of 
Plant Diseases and Disorders in Indiana — 1974. Proc. Indiana Acad. Sci. 
84:71-84. 

3. Wolf, S. C. 1972. Plant Diseases in Indiana in 1972. Proc. Indiana Acad. Sci. 
82:101-108. 



Chromium-Zinc Interaction in Accumulation of 
Minerals by Bush Beans 

Heidi A. Schmitt and Connie M. Weaver 

Foods and Nutrition Department 

Purdue University, West Lafayette, Indiana 47907 

Introduction 

Zinc is necessary for human metabolism. It is a necessary trace element in 
plants for the synthesis of tryptophan and anxin and also functions in metaloen- 
zyme systems. Chromium has been implicated as being necessary for efficient car- 
bohydrate metabolism and has been shown to be effective in treating other dietry 
related diseases. Chromium is not considered to be an essential element for 
plants, but no study has been able to completely eliminate the element from the 
plant. Huffman and Allaway (4) attempted to eliminate chromium from plants (ro- 
maine lettuce, tomato, wheat, and bush bean) by growing successive generations 
in "chromium-free" solution (3.8xl0~ 4 fiM Cr) and reduced the chromium levels to 
an average of 22 ng Cr/g dry wt. Chromium may be necessary in very minute 
quantities, and it has been shown that small quantities of chromium in the 
nutrient solution enhance growth of avacodo and citrus plants compared to a 
"chromium-free" solution (2). 

Hahn and Evans (3) reported an antagonism between zinc and chromium in 
the absorption of these metals by rats. A chromium and zinc interaction in ac- 
cumulation of these minerals in plants has not been reported. In this work, bush 
beans were choosen as a model for investigation of a possible chromium-zinc in- 
teraction. 

Methods 

Bush beans {Phaseolus vulgaris L. 'Blue Lake') were germinated in a cir- 
culating hydroponic system using BR-8 cellulose grow blocks as supports. Seven 
days after germination each group of 15 plants was exposed to a nutrient solution 
(1) containing the following: 2.2xl0 5 cpm 51 Cr/liter, 8.8xl0 5 cpm 65 Zn/liter, or both 
nuclides dosed at the same concentration as in the single labeled solutions. All 
solutions contained 1.5 ppm stable Zn and no added stable chromium. In the 
radioactive solutions, 2.2xl0 5 cpm 51 Cr/liter represents less than 3xl0' 2 ppb and 
8.8xl0 5 cpm 65 Zn/liter represents less than 1.02 ppb. 

The beans were harvested at maturity (2 months after germination), washed 
with deionized water, and chopped to a very fine mesh in a food processor. 
Samples (approx. 9g. wet wt.) were taken from each group and dried in a vacuum 
over at 70°C for 12 hours. Samples were then counted using a Harshaw 3" x 3" 
NaKTl) integral line detector. Overlapping of the energy spectrum of chromium 
and zinc in the dual-labeled plants was accounted for by solving simultaneous 
equations. Counts were adjusted for fluctuations in counting efficiency and for 
decay occuring after harvest. 

Results and Discussion 

Accumulation of 51 Cr by bush beans in the presence and absence of 65 Zn is 
shown in Figure 1. Bush beans exposed only to radioactive chromium contained 
303 ± 17 cpm/g dry wt. which was 76.9% of the count rate (398 ± 20 cpm 51 Cr/g 
dry wt.) accumulated by the dual-labeled plants. 

125 



126 



Indiana Academy of Science 






o 



E 

a 



2 



1 



Cr Cr/Zn 

Treatment 

FIGURE 1. 51 Cr accumulation in the absence and presence of 65 Zn in bush beans. 



The influence of 51 Cr in the nutrient solution on accumulation of 65 Zn by bush 
beans is shown in Figure 2. Beans exposed to radioactive zinc alone accumulated 
8025 ± 90 cpm 65 Zn/g dry wt. while those grown in nutrient solutions containing 
both radionuclides accumulated 1358 ± 37 cpm 65 Zn/g dry wt. This represents an 
accumulation by dual-labeled plants of only 16.9% as much 65 Zn as single-labeled 
plants. 

These results show that the accumulation of chromium by plants is somewhat 
increased by zinc, the accumulation of zinc is greatly suppressed by chromium at 
least for the levels used in this study. Two possible explanations of these observa- 
tions can be offered. Competition between chromium and zinc at the site of uptake 
of these elements or for translocation into the bean may exist. This explanation is 
unlikely since the presence of radioactive zinc did not suppress the uptake of 
radioactive chromium. Alternatively, the level of chromium used in this study 
may have been toxic to the uptake or translocation mechanisms for zinc which 
could influence zinc accumulation by the plants. Toxicity due to chromium has 
been noted at levels of > 1.0 ppm in soybeans (8) and these authors suggested that 
chromium may exert its toxic effects on the roots. Zinc is known to be readily 
translocated in plants and its concentration in the shoots varies linearly with the 



Botany 



127 



8 



7- 



* 5 

l. 
■o 

d) 4 



K 

E 

a 

w 

m 

o 

i 
c 

N 



3 



Zn 



Zn/Cr 



Treatment 

FIGURE 2. 65 Zn accumulation in the absence and presence of 51 Cr in bush beans. 



128 Indiana Academy of Science 

dose (9), chromium is poorly translocated and most chromium resides in the roots. 
An increase in the chromium concentration in the nutrient solution does not 
necessarily mean an increase in chromium concentration in the plant (7). Others 
have shown the beneficial effects of chromium to be within a very narrow concen- 
tration range (2) which may vary with the concentration of other macrominerals in 
plants (6) and between plant species. 

Plants treated with 51 Cr showed a decreased yield. The plants also exhibited 
signs of early senescence and wilting and browning of roots compared to plants 
grown in the absence of 51 Cr. Similar symptoms have been noted in chromium tox- 
icity studies (5). 

Although the yields were not recorded in this experiment, as part of another 
study in which bush bean plants were exposed to 65 Zn and to the dual label of 65 Zn 
and 51 Cr, the yield was calculated for three plants from each treatment. The bean 
yield from the dual-labeled plants was only 41% of the yield of single-labeled 
plants. A similar suppression of zinc accumulation by chromium was observed in 
these plants. 



Literature Cited 

Furuya, M. and W. S. Hillman. 1964. Observations in spectrophotometri- 
cally assayable phytochrome in vivo in etiolatic pisum seedlings. Plant 
Physiol. 63:31. 

Haas, A. R. C. and J. N. Brusca. 1961. Effects of chromium on citrus and 
avocado grown in nutrient solutions. Calif. Agri. 15(2):10. 
Hahn, C. J. and G. W. Evans. 1975. Absorption of trace metals in zinc- 
deficient rat. Am. J. Physiol. 228(4):1020. 

Huffman, E. W. D. and W. H. Allaway. 1973. Growth of plants in solution 
culture containing low levels of chromium. Plant Physiol. 52:12. 
Hunter, J. G. and 0. Vergnano. 1953. Trace elements toxicities in oat 
plants. Annuals of Applied Biology. 40:761. 

Soane, B. D. and Saunders, D. H. 1959. Nickel and chromium toxicity of 
serpentine sails in southern Rhodesia. Soil Science. 88:322. 
Thomas, B., J. A. Raughan, and E. D. Walters. 1974 Cobalt, chromium, and 
nickel of some vegetable foodstuff. J. Sci. Fd. Agri. 25:771. 
Turner, M. A. and R. H. Rust. 1971. Effects of chromium on growth and 
mineral nutrition of soybeans. Soil Science Soc. Amer. Proc. 35:755. 
Weaver, C. M. and H. A. Schmitt. 1980. Unpublished data. 



CELL BIOLOGY 

Chairman: Stanley N. Grove 
Department of Biology, Goshen College, Goshen, Indiana 46526 

Chairman-Elect: Kara W. Eberly 
Department of Biology, St. Mary's College, Notre Dame, Indiana 46556 



ABSTRACTS 

Observations on the Tertiary Structure of Ribosomes Released from Membranes 
of Rough Endoplasmic Reticulum. John H. Elder and D. James Morre, Purdue 

University, West Lafayette, Indiana 47907. Much knowledge concerning the 

tertiary structure of ribosomes has been deduced from morphological and 
biochemical studies in which ribosomes have been examined by electron 
microscopy either from thin sections or in negatively stained preparations or from 
studies where ribosomes have been dissociated into subunits and component pro- 
teins and reassembled as well as from studies of attachment of mRNA and the 
growing polypeptide chain. Generally, ribosomes consist of a major and minor 
subunit with a groove or channel to accomodate the mRNA plus a channel in the 
large subunit to direct the growing peptide chain out of the ribosome and into the 
membrane lumen or soluble cytoplasm. In the present study, image enhancement 
of negatively stained ribosomes from polyribosomes released from rough endo- 
plasmic reticulum of rat liver has been used to provide a 3-dimensional view of one 
class of eukaryotic ribosomes. Work supported in part by a grant from the Na- 
tional Science Foundation. 

Complex Gangliosides Bind Fibronectin. David C. Evers and D. James Morre 

Purdue University, West Lafayette, Indiana 47907. Surface properties of cells 

that permit tumors to metastasize are poorly understood. In normal cells, 
fibronectin, a large glycoprotein, may be employed as a linker molecule permitting 
cells to attach to a collagen-rich stroma. The present study identifies several com- 
plex gangliosides that bind fibronectin and may serve as the cellular binding site 
for fibronectins. Findings will be correlated with results of previous studies from 
our laboratory where ganglioside levels have been determined and compared in 
metastatic and non-metastatic hepatocellular carcinomas of the rat. Work sup- 
ported in part by a grant from the American Cancer Society. 

Electrophoretic Study of the Deoxyribonuclease Activity of Human Urine. D. K. 

Hansen, W. Chaekal and M. E. Hodes, Department of Medical Genetics, Indiana 
University School of Medicine, Indianapolis, Indiana 46223. The deoxyribonu- 
clease activity present in human urine was analyzed by several common elec- 
trophoretic techniques. Multiple bands of activity can be resolved following elec- 
trophoresis in polyacrylamide gels, incubation of the gel with an agarose overlay 
containing DNA, precipitation of the undigested DNA and staining of the DNA 
with methyl green or toluidine blue. After isoelectric focusing, bands of activity 
with wide differences in pH were visualized. The focusing pattern was not 
affected by treatment with neuraminidase. Common disc electrophoretic systems 
were used to study these activities. Smearing of some bands made analysis of 
anodally migrating activity difficult. The cathodally migrating bands from over 
100 urine samples have been analyzed. The relative mobilities of the resultant one 

129 



130 Indiana Academy of Science 

or two bands generally seen are quite reproducible. Occasionally a third band is 
visible closer to the origin. Treatment of the urine with neuraminidase prior to 
electrophoresis has no effect on the band number or mobility. In addition, analysis 
of low (to 50 pg) quantities of bovine pancreas DNase in the pH 4.5 cathodal 
system did not reveal the variation in mobility with amount of enzyme applied 
that had previously been demonstrated for RNase A and human urine RNase. It is 
thought that this invariance of mobility is due to the lack of anion binding by the 
relatively acidic (pi 4.7) DNase I as opposed to the extreme anion binding by the 
very cationic (pi ca. 9.5) RNase. 

Characterization of A Fast Transported Calicum — Binding Protein in Mammalian 
Nerve. Zafar Iqbal, Departments of Physiology, Biochemistry and Medical 
Biophysics, Indiana University School of Medicine, Indianapolis, Indiana 46223. 

Axoplasmic transport, a process required to carry down cellular components 

essential for the maintenance of proper form and function of nerve, was found depen- 
dent on Ca 2+ . Following an injection of 45 Ca 2+ into the L-7 dorsal root ganglion of 
cat, 45 Ca 2+ radioactivity was found to be associated with a calcium-binding pro- 
tein (CaBP) of about 15,000 dalton and both were transported in sciatic nerve at a 
fast rate close to 410 mm/day (Iqbal and Ochs, 1978; J. Neurochem. ,17:409-418). 
This CaBP now has been isolated from dog sciatic nerve and purified by a com- 
bination of separation techniques such as gel filtration on Sephadex G-100 and 
G-25, ion exchange chromatography on DEAE-cellulose and preparative isoelec- 
trofocusing. The purified CaBP shows similarities to calmodulin, a calcium- 
dependent regulatory protein in regards to its mobility on SDS containing 
polyacrylamide gels, and analytical isoelectro-focusing gels and activation of cyclic 
nucleotide phosphodiesterase. A fast transport of CaBP in the nerve and its 
similarities to calmodulin suggests its involvement in the axoplasmic transport of 
materials in nerve fibers. In the transport filament model proposed for fast ax- 
oplasmic transport, energy supplied by the hydrolysis of ATP by Ca-Mg-ATPase 
is required to move the transport filament to which various components are 
bound, down the microtubules. The activity of Ca-Mg-ATPase and the assembly- 
disassembly of microtubules has been shown to be regulated by calmodulin. Addi- 
tionally, calmodulin regulates a number of enzymes including those involved in 
the metabolism of carbohydrates required to maintain the proper supply of ATP 
needed for the transport system to operate. Supported by NIH ROl 8706-11, 
American Cancer Society In-46T and the Indiana Academy of Science. 

The Role of Beta-alanine in Pigmentation Polymorphisms. M. E. Jacobs, Depart 

ment of Biology, Goshen College, Goshen, Indiana 46526. The influence of 

beta-alanine in integumentary pigmentation is investigated. In addition to its 
presence in tanned insect cuticles, beta-alanine is present in the tanned myeclial 
walls of the fungus, Morchella esculenta and in human hair. Its possible role in 
pigmentation of such structures is discussed. Beta-alanine forms yellow-red tan- 
ning pigments when incubated with N-acetyldopamine or leucyltyrosine oxidized 
by tyrosinase, or with unsaturated phospholipids, such as lecithin, oxidized by 
heat. 

Antioxidants, Fatty Acids and Oxidant Stress, and the Control of Cell Prolifera- 
tion in Culture. James S. Miller, Division of Natural Sciences, Goshen College, 
Goshen, Indiana 46526 and Samuel 0. Ikharebha, Victor C. Gavino, George E. 
Milo, and David G. Cornwell, Ohio State University, Columbus, Ohio 43212. 

Previous work showed that several polyunsaturated fatty acides inhibit the 

proliferation of smooth muscle cells and fibroblasts in tissue culture, a-tocopherol 



Cell Biology 131 

(Vitamin E) and other antioxidants had the opposite effect, enhancing prolifera- 
tion of smooth muscle cells. 

In the present study we show that polyunsaturated fatty acids produce oxi- 
dant stress in smooth muscle cells, as measured by the thiobarbituric acid (TBA) 
assay for lipid peroxidation, a-tocopherol was an inhibitor of this lipid peroxida- 
tion produced by polyunsaturated fatty acids. 

a-tocopheryl quinone (Vitamin E-quinone), a metabolite of a-tocopherol, was 
better than a-tocopherol as a stimulator of the proliferation of both smooth muscle 
cells and fibroblasts in culture, a-tocopherylquinone was also better than 
a-tocopherol in inhibiting the formation of lipid peroxides in cells incubated with 
polyunsaturated fatty acids. These results demonstrate that a-tocopherylquinone 
is an antioxidant in vivo. 

Several other quinones also were studied: menadione, phylloquinone, and 
ubiquinone-10. Menadione was a potent inhibitor of smooth muscle cell prolifera- 
tion in culture; the other two quinones had no effect. The different results with 
these quinones may be due to specific structural differences. 

Effects of Cytochalasins on Selected Species of Aspergillus and Mucor. Kenneth 

C. Miller and Stanley N. Grove. Goshen College, Goshen, Indiana 46526. The 

effects of cytochalasins are surveyed on a variety of Aspergillus species. The 
intent is to clarify the relationship between sensitivity to the inhibitors and poten- 
tial pathogenicity. Likewise, several Mucor species are similarly surveyed to test 
the generalization that fungi which are capable of causing infections of deep organs 
in higher animals are characterized by an ability to develop in the yeast form. The 
sensitivity to the cytochalasins of dimorphic species is compared with that of non- 
dimorphic species. Our preliminary findings suggest that some pathogenic or 
dimorphic forms are less sensitive to cytochalasins than closely related non- 
pathogenic or non-dimorphic forms. Supported in part by a grant from the National 
Institute of Allergy and Infectious Diseases. 

Effect of Vitamin A on the Heptaic Golgi Apparatus Architecture. Dorothy M. 
Morre, D. James Morre and Martha Walter, Department of Foods and Nutri- 
tion and Department of Medicinal Chemistry and Pharmacognosy, Purdue Univer- 
sity, West Lafayette, Indiana 47907. The role of the Golgi apparatus in glycosy- 

lation was shown first by autoradiography with goblet cells, which secrete large 
quantities of mucin, a mucoprotein of high sugar content. The sulfation process, 
that is the transfer of sulfate from an activated donor to an appropriate receptor, 
requires the activity of sulfotransferases and may occur in the Golgi apparatus as 
well. Vitamin A is required not only for activation of sulphatide but also for other 
processes of mucopolysaccharide synthesis. In this study, we investigated the 
effects of vitamin A on the architecture of the Golgi apparatus. Male weanling CDF 
rats were fed diets containing either no, adquate (4,000 IU/kg diet) or excess (100 
times adequate) amounts of vitamin A as retinyl acetate. Animals were killed at 2 
to 8 weeks postweanling and portions of the liver were prepared for electron 
microscopy. Quantitative and qualitative determinations were made from mont- 
ages of randomly selected whole parenchymal cells. The extent of individual Golgi 
apparatus stacks (dictyosomes) paralleled vitamin A intake. Relative to controls, 
there was a reduction in total membrane surface occupied by Golgi apparatus mem- 
branes in cells of vitamin A deficient livers while there was an increase in excess. 
Secretory vesicles and vesicular profiles appeared unaffected by vitamin A 



132 Indiana Academy of Science 

extremes. Generally, Golgi apparatus of livers of vitamin A deficient animals 
appeared smaller with more fenestrated cisternae while livers of excess animals 
had more extensive sheet-like cisternae with more well-developed complex cister- 
nae on the mature face in the GERL region. The overall appearance of the Golgi ap- 
paratus architecture was influenced by dietary vitamin A levels. Supported by 
grants from the NIH CA 18801 and the American Cancer Society. 

Distribution of Acid Phosphatase in Transplanted Hepatomas of the Rat. Dorothy 
M. Morre and Ann L. Rosenthal, Department of Foods and Nutrition, Purdue 

University, West Lafayette, Indiana 47907. Acid phosphatase is considered to 

be a marker enzyme for lysosomes even though it is present in other parts of the 
cell (e.g. Golgi apparatus). In our work, we have combined determinations of acid 
phosphatase using /^-glycerophosphate as a substrate with simple cell fractionation 
procedures. Male weanling inbred CDF rats were subcutaneously injected with 
hepatocellular carcinoma cells of the rat derived from solid tumors induced in 
syngeneic donors by 2-acetylaminofluorene. Livers were homogenized and frac- 
tionated into nuclear, mitochondrial-lysosomal and microsomal fractions by dif- 
ferential centrifugation. Total activity of acid phosphatase was measured as acti- 
vity present when Triton X-100 was added to the incubation medium whereas free 
activity was the activity present with no added detergent. Fraction composition 
was confirmed by morphometric measurements determined from electron 
micrographs. Both total and latent acid phosphatase activities in all hepatoma frac- 
tions were significantly less than corresponding fractions from control livers. 
Based on latent activity, the lysosome populations would be about V2 that of normal 
liver. 

Supported by The American Cancer Society and the Purdue Agricultural Ex- 
periment Station. 

Exocytosis: Routes and Kinetics of Delivery of Secretory and Membrane Pro- 
teins. D. James Morre, Edward M. Croze, Cheryl Franz and C. M. Eppler. Pur- 
due University, West Lafayette, Indiana 47907. Vectorial movement (flow) of 

membranes is an integral part of the activities of most eukaryotic cells. Involved 
are various endomembrane components including endoplasmic reticulum (ER), 
Golgi apparatus (GA) and plasma membrane (PM) as well as vesicles and tubules 
associated with each of the major structures. Studies that correlate movement of 
both membrane constituents and constituents of secretaory products 
simultaneously in the same experimental system have been few. Our studies with 
rodent liver, correlate exocytosis of membrane markers, mouse H-2 histom- 
patibility antigens and nucleoside diphosphate phosphatase, with exocytosis of 
secretory lipoproteins and albumin. A correlative approach utilizes metabolically 
labeled constitutents, highly purified cell fractions, and specific immunoprecipita- 
tion analyses. The order of labeling is ER, GA and PM; turnover kinetics suggest 
transfer of both membrane and secretory product from ER to GA with accumula- 
tion in PM or the extracellular environment. Work supported by grants from the 
National Institutes of Health and the National Science Foundation. 

Comparative Cytochemical Studies of Onion Root Tip with Specific and Non- 
specific Substrates for Nucleoside Diphosphatase(s). Michael J. Stewart and 
Charles W. Goff, Department of Life Sciences, Indiana State University, Terre 

Haute, Indiana 47809. Nucleoside diphosphatase (NDPase) has been used as a 

marker enzyme for the Golgi apparatus in a number of plant and animal systems. 



Cell Biology 133 

Previous studies from our laboratory have shown (1) NDPase to be localized in a 
number of subcellular sites and (2) that there are two classes of NDPase 
isoenzymes-one which hydrolyzes only nucleoside diphosphate substrates and 
another class which not only hydrolyzes nucleoside diphosphates but also certain 
nucleoside monophosphates and thiamine pyrophosphate (TPP). Cytochemical 
studies at neutral pH were undertaken to determine whether different subcellular 
localization patterns could be observed using different nucleotides and TPP, 
which are hydrolyzed by one or both classes of onion root tip NDPases. 

IDPase reactivity occurred in the Golgi apparatus, endoplasmic reticulum, 
nuclear envelope, plasmalemma, cell wall, and tonoplast of certain tissues of the 
onion root tip. ADPase reactivity was found in the ER, nuclear envelope, 
tonoplast, and cell wall of various root tissues although the intensity of staining 
was less than that seen for IDPase. Very little ADPase reactivity was found in the 
Golgi apparatus and very little, if any, reactivity was found on the plasmalemma. 
TPPase reactivity was similar to that seen using ADP except that there was very 
intense reaction product in the definitive cell wall surrounding lateral cap, epider- 
mal, and cortical cells. Little, if any, reaction product appeared in the Golgi 
apparatus in tips incubated with TPP. AMPase reactivity was found in the ER, 
cell wall, and vacuoles of certain tissues and the staining was much less intense 
relative to IDPase reactivity. No reaction product was found in the Golgi 
apparatus. Preliminary results from gel filtration studies indicate that peaks of 
enzyme activity may be shared by enzymes which hydrolyze IDP, other 
nucleotides, and TPP. 

Our results lend additional support to previous findings which suggest that 
there are two classes of NDPase isoenzymes: one which may be referred to as 
specific NDPases, the other as non-specific phosphatases. The findings from our 
present investigation suggest the potentiality for discriminating between these 
two classes of NDPases with the ultimate goal of clarifying the marker enzyme 
question for the onion root tip. 

Cytochalasin A (0.1-1 ug/mll Rapidly Halts Hyphal Tip Growth in Rhizoctonia 
solani. James A. Sweigard and Stanley N. Grove, Goshen College, Goshen, 

Indiana 46526. Within two minutes of inhibitor application growth slows, 

Spitzenkorper integrity is disrupted and hyphal tips become bulbous. Our initial 
observations suggest that the phase-light area of the Spitzenkorper immediately 
retracts from the hyphal apex upon treatment but that growth continues for a 
short time. A possible mechanism for this inhibition is the disruption of essential 
microfilaments in the tip region. Supported in part by a grant of the National 
Institute of Allergy and Infectious Diseases. 

Kinetics of Appearance of Lipoprotein Particles in Cisternae of Subsurface 
Smooth Endoplasmic Reticulum of Isolated Rat Livers Perfused with Free Fatty 
Acid. Martha Twaddle, Ralph A. Jersild and James Morre, Indiana Univer- 
sity School of Medicine, Indianapolis, Indiana 46223, and Purdue University, West 

Lafayette, Indiana 47907. Vesicles containing single lipoprotein particles are 

found in the pericapillary cytoplasm of rat hepatocytes in the proximity of particle 
containing rough-smooth endoplasmic reticulum transition elements. These 
lipoprotein-containing smooth endoplasmic reticulum elements provide an alter- 
native source to Golgi apparatus as a supplier of very low density lipoprotein par- 
ticles to the circulation. To test this possibility, the kinetics of appearance of 
lipoprotein particles in these elements was determined for isolated rat livers per- 



134 Indiana Academy of Science 

fused with free fatty acid. The results show that the smooth endoplasmic 
reticulum elements of the pericapillary cytoplasm acquire lipoprotein particles in 
advance of elements of the conventional Golgi apparatus and that the particles 
may be secreted directly to the circulation in a manner completely bypassing the 
Golgi apparatus. 

Persistent Nucleoli in Various Meristems of Phaseolus aureus (mung bean). 

Martin A. Vaughan, Department of Life Sciences, Indiana State University, 
Terre Haute, Indiana 47809 and J. P. Braselton, Department of Botany, Ohio 
University, Athens, Ohio 45701. The nucleolus, a non-membrane-bound struc- 
ture characteristic of interphase nuclei of eukaryotic cells, is the site of ribosomal 
ribonucleic acid (rRNA) synthesis. Nucleoli generally do not arise from pre- 
existing nucleoli by division, but undergo cyclic changes during the mitotic cycle. 

Persistence of the nucleolus during mitosis in vascular plants is not an 
unusual phenomenon and in many groups persistence of the nucleolus to 
metaphase and beyond is common. In addition to their natural occurrence, persis- 
tent nucleoli also may be induced by experimental conditions, such as exposure to 
cobalt salts, 5-fluoro-deoxyuridine (FUdR), ethidium bromide, relatively low 
temperatures, and centrifugation. 

Most observations of persistent nucleoli in higher plants were described from 
cell divisions in root tips of recently germinated seedlings; and a few cases were 
reported in root tips of mature plants. Peristent nucleoli at metaphase in primary 
root, secondary root, and shoot apical meristems of the mung bean were observed 
by light microscopy of hematoxylin-stained paraffin sections 4, 8, and 14 days 
after germination. The percentage of metaphases containing persistent nucleoli 
were recorded in each meristem to establish whether persistence of nucleoli at 
metaphase is associated with the age or the type of meristem. 

There was a significantly lower percent of persistent nucleoli at metaphase 
in shoot apical meristems as compared to primary or secondary root meristems at 
each of the ages tested. There was no significant difference in percent of persis- 
tent nucleoli at metaphase in the primary or secondary root meristems. 

The variation in nucleolar behavior of mung bean shoot and root meristems 
may be explained by differences in the length of the nucleolar dispersion cycle or 
differences in the length of the mitotic prophase corresponding to the dispersive 
phase of the nucleolar cycle. 

The Effect of Sympathectomy on the Structure of the Rat Pineal Gland. Henry C. 
Womack and Mohinder S. Jarial Department of Physiology and Health Science 
and Muncie Center for Medical Education, Ball State University, Muncie, Indiana 

47306. Female albino rats at the age of 4 months were bilaterally sympathec- 

tomized under ether anesthesia by surgical excision of a small segment of the 
sympathetic trunk just below the superior cervical ganglion to disrupt the light 
pathway to the pineal gland. Control animals were subjected to a sham operation. 
The appearance of ptosis in the experimental animals provided confirmation of 
complete sectioning of the sympathetic trunk. 

Following surgery the animals were maintained under conditions of 14 hours 
light and 10 hours darkness with food and water ad libitum. After 7 days the 
animals were sacrificed under either anesthesia by intravascular perfusion with 
2% glutaraldehyde. The pineal was removed and fixed in a mixture of 
glutaraldehyde and osmic acid in phosphate buffer for electron microscopy and in 
Bouin's fluid for light microscopy. 



Cell Biology 135 

Electron microscopy revealed small membrane bound granules in the vicinity 
of endoplasmic reticulum and Golgi apparatus. In the experimental animals the 
number of such granules was enhanced as compared to the controls. There was no 
difference in the number of lipid droplets in the experimental and cntrol animals. 
Epon 1 micron thick sections stained with azur II, showed medium to large size 
granules which in thin sections stained with uranyl acetate and lead citrate, 
revealed an outer concentric ring. Such granules were present in relatively larger 
numbers in the experimental animals than in the controls. Occasionally these 
granules were seen in the lumina of capillaries. It appears that these large 
granules in rats represent concretions or brain sand which in humans have been 
called corpora arenacea. Numerous nerve terminals were observed in the 
perivascular space and some in association with pinealocytes. In the experimental 
animals the nerve terminals contained empty vesicles while in the controls 
vesicles were granulated. 

The results of this study suggest that in the absence of light input to the 
pineal gland as a result of the lesion of preganglionic sympathetic fibers, the 
postganglionic neurons and nerve terminals fail to synthesize neurotransmitters 
such as norepinephrine and serotonin. It appears that the secretory activity of the 
pinealocytes is not affected by sympathectomy, but the secretion products are not 
released for use by the animal. In the control animals secretion products that are 
being synthesized are also being utilized. It is suggested that some of the 
secretory material like melatonin produced by the pinealocytes may be 
accumulated in the form of hard granules or concretions which have been observ- 
ed in large numbers in the experimental animals. 

Supported by an academic year 
grant from Ball State University. 



Effect of Maternal Thyroid Activity Upon 
In Vitro Protein Synthesis in the Rat Blastocyst 

Vernon G. Archer, Department of Biology 
Jackson State Univerity, Jackson, Mississippi 39203 

and 

James P. Holland, Department of Biology 

Indiana University, Bloomington, Indiana 47405 

Introduction 

Many investigators have reported that thyroid hormones influence the 
reproductive system and pregnancy. The literature contains conflicting reports 
concerning the extent and effect of this influence. Examples of these conflicts and 
possible explanations were presented in an earlier publication (10) and have been 
reviewed by Leatham (13). Despite numerous investigations into this 
phenomenon, the mechanism by means of which thyroid hormone exerts its effect 
upon reproduction and pregnancy remains unresolved. 

In our laboratory, earlier studies of this phenomenon utilized the technique 
of experimentally delayed implantation of blastocysts (6) which allows the investi- 
gator to control some of the variables in studies of reproduction. Using this techni- 
que, it was demonstrated (10 and 12) that in rats receiving daily injections of 
either 48 ,ug or 8 fig L-thyroxine, the hyperthyroid condition was able to signifi- 
cantly counteract the detrimental effects of progesterone deficiency upon the 
number of surviving blastocysts on Day 8 of pregnancy or the number of implant- 
ing blastocysts on Day 13 of pregnancy. Surgical thyroidectomy had the opposite 
effect. The thyroid effect was shown to be exerted during the progesterone- 
dependent delay period rather than during the estrogen-dependent implantation 
process. Also, our earlier work (11) showed that the sub-normal levels of uterine 
alkaline phosphatase which occur during progesterone deficiency were restored to 
normal by thyroxine treatment, but were further depleted by a hypothyroid con- 
dition in rats examined at a time corresponding to Day 8 of pregnancy (the final 
day of the progesterone-dependent delay period). It was further demonstrated 
that the thyroxine effect on uterine alkaline phosphatase was not due to a 
generalized action on metabolism. Thus, thyroid hormone does exert an influence 
upon blastocyst survival by means of a modification of progesterone-dependent 
uterine physiology, but the possibility also exists that thyroid hormone may exert 
a more direct influence upon the blastocyst itself. The present investigation was 
designed to study this possibility by examining protein and nucleic acid 
metabolism of blastocysts during alterations of the progesterone and thyroid hor- 
mone levels. 

Materials and Methods 

Sprague-Dawley-derived female albino rats, between 60 and 100 days old 
were maintained on Purina Laboratory Chow and tap water ad libitum. Rats were 
housed in an animal room at 24 °C with a daily illumination schedule of 14 hours of 
light and 10 hours of darkness. Females showing a proestrus or early oestrus 
vaginal smear were placed overnight in cages with adult male rats. Insemination 
was confirmed on the following morning by the presence of spermatozoa in the 
vaginal smear and this was designated as Day 1 of pregnancy. Hyperthyroidism 
was induced by daily injection of 12 fig L-thyroxine beginning at least ten days 

136 



Cell Biology 137 

prior to insemination. Surgical thyroidectomies were performed through a 
midventral incision in the neck at least four weeks prior to insemination. 

Blastocyst Collection and Incubation 

Blastocysts were collected from control, hyperthyroid and hypothyroid 
animals which were sacrificed on Day 5 of pregnancy. The uterus was excised, the 
cornua were severed from the vagina, and each horn was flushed using a 23 guage 
hypodermic needled fitted to a one-ml syringe containing Basic Medium for Ova 
Culture-2 (2). The blastocysts were flushed into depression slides using a minimal 
volume of BMOC-2. Finely drawn Pasteur pipettes were then used to place the 
blastocysts in culture dishes for incubation in BMOC-2 containing the radioactive 
precursors. The BMOC-2 was previously sterilized by filtration through a 
millipore filter. 

Approximately 0.5 ml of culture medium was placed in a 60 x 15 style 3010 
plastic organ tissue culture dish (Falcon Plastics, Oxnard, Calif.), equipped with an 
absorbent ring which was saturated with water to provide the inside of the dish 
with water vapor. Uniformly labeled 14 C-leucine (specific activity between 1 and 
3mCi/mM) was added to the BMOC-2 to give a concentration of 1 x 10 5 mM. Be- 
tween 10 and 20 blastocysts were placed in each dish and the dishes were placed 
for 1.5 hours in a Model 3212 water-jacketed incubator (National Incubator) at 
37°C with a C0 2 -0 2 atmosphere. To verify protein synthesis, incubations were 
done in both the presence and absence of cycloheximide (5 /xg/ml or 50 figlml). 

14 C-Amino Acid Uptake and Incorporation 

At the end of the incubation period, the blastocysts were collected in approx- 
imately 5 fil of medium and rapidly washed by means of sequential transfers 
through three changes of two ml of BMOC-2 containing non-radioactive leucine. 
The blastocysts were then transferred in approximately 5 ^1 of the final wash to a 
2 ml centrifuge tube containing 100 fil of water to cause cellular lysis. Using dry 
ice and acetone, the water containing the blastocysts was frozen and thawed three 
times, and was agitated on a vortex mixer during each thawing. One mg of bovine 
serum albumin and 1 mg of glycogen in 20 nl of water were added to each tube to 
act as cold carrier and thus reduce the loss of labeled compounds during the ex- 
perimental manipulations (3). 

The protein from the blastocysts was precipitated, extracted and prepared 
for counting according to the method reported by Brinster (3) with the exception 
that Aquasol was used as the scintillation fluid. This procedure involved precipita- 
tion of the protein with 5% trichloroacetic acid (TCA) and counting the TCA- 
washed precipitate (acid insoluble) as the amount of the 14 C-leucine which was in- 
corporated into protein. An aliquot of the TCA supernatant (acid soluble) which 
had been drawn off and measured also was counted and used to calculate the 
radioactivity of the entire supernatant or acid soluble fraction. The sum of the 
14 C-radioactivity in the acid insoluble (incorporated) and acid soluble fractions was 
considered to be the total 14 C uptake (total uptake = incorporated + acid 
soluble). All counts of radio-activity were made using a Model LS-233 liquid scin- 
tillation counter. 

Autoradiography 

Delayed blastocysts were obtained as described by Holland et al. (12) involv- 
ing ovariectomy on Day 3, daily injection of progesterone (2.0 mg/day or 0.4 mg/- 
day), and flushing of the uteri on Day 8 of pregnancy. The blastocysts were in- 



138 Indiana Academy of Science 

cubated using the same procedure as described above with the exception that the 
radioactive precursors which were added to the BMOC-2 (at a concentration of 1 x 
10" 4 mM) were 3 H-uridine(6 uCi/ml, 20-25 (Ci/mole) or 3 H-phenylalanine (6 uCi/ml, 
170-200 mCi/mM) for a five hour incubation period. Studies were also done using a 
concentration of 1 fiM of 3 H-uridine in the incubation medium since this has been 
shown (7) to be a saturation level which eliminates differences in incorporation 
which may be associated with the concentration of precursor in the medium. Some 
incubations were performed in the presence of actinomycin D (0.1 /-tg/ml) or 
cyclohexomide (5 uglml or 50 /ig/ml). After incubation the blastocysts were remov- 
ed and washed as described earlier. Several blastocysts from each treatment 
group were placed on separate microscope slides which had been subdued with 
albumen and were prepared for autoradiography in a manner similar to that 
described by Skalko (16) using Carnoy's fixative. Blastocysts were treated with 
2% perchloric acid at 4°C for 20 minutes to remove free nucleotides and some 
slides were subjected to ribonuclease extraction (1 mg/ml at 37°C for 2 hours) to 
confirm the specificity of incorporation of the uridine. Slides were coated with a 
1:1 dilution of Kodak Nuclear Emulsion (NTB-3, exposed for seven days), 
developed in Kodak D-19 for four minutes at 18°C, acid fixed, and stained with 
hematoxylin and eosin. 

Results 

Table 1 summarizes the data from in vitro studies concerning 14 C-leucine up- 
take and incorporation into protein by blastocysts which had been flushed from 
uteri of thyroidally altered rats on Day 5 of pregnancy. When compared with 
blastocysts from control (euthyroid) rats, the blastocysts from both hyperthyroid 
and from hypothyroid rats showed a significantly greater uptake and incorpora- 
tion of 14 C-leucine. However, the percent of the 14 C-leucine uptake which had been 
incorporated into protein by these blastocysts was approximately twice the con- 
trol level in blastocycsts from hyperthyroid rats and was only about one-half the 
control level in blastocysts from hypothyroid rats. These differences were 
statistically significant. Since there was low incorporation of the amino acid in the 
blastocysts from hypothyroid animals, the disproportionally high uptake is 
possibly associated with extracellular movement of fluids which occurs during 
hypothyroidism. In this case, possibly there is movement of fluid (containing 
radioactive leucine) into the cavity of the blastocyst and thus a disproportionally 
high uptake is registered for these blastocysts. 



Table 1. In Vitro Uptake and Incorporation of U C Leucine by Blastocysts From 
Thyroidally Altered Rats 



Thyroid State Total Incorporation % Incorporation of 

Uptake Uptake into Protein 

(cpm/blastocyst) cpm/blastocyst) 

Euthyroid 24.40 ±3.50 6.42 ±0.28 26.3 ±4.1 

Hyperthyroid 43.12 ±7.50* 27.62 ±3.78* 64.1 ± 8.3* 

Hypothyroid 103.64 ±8.95* 13.90 ±2.88* 13.4 ±2.2* 

♦Significant at the 5% level by "t" test 

(Each thyroid state represents results from five groups consisting of 15-20 blastocysts/group.) 



Cell Biology 139 

Table 2 summarizes the data from the autoradiography of experimentally 
delayed blastocysts which had been incubated for five hours in BMOC-2 contain- 
ing 3 H-phenylalanine or 3 H-uridine. Blastocysts from euthyroid rats showed essen- 
tially identical amounts and distribution of the precursors regardless of whether 
these rats were receiving a low or normal maintenance dose of progesterone. 
Blastocysts from hyperthyroid rats clearly showed increased amounts of both 
precursors in the inner cell mass (embryonic disc). The changes in the trophoblast 
area were less clearly defined but tended to show a slightly decreased concentra- 
tion. Blastocysts from thyroidectomized rats of both progesterone treatment 
levels generally showed decreased incorporation of precursors in the trophoblast 
but those blastocysts from the thyroidectomized rats which had been maintained 
on a deficiency dosage of progesterone (0.4 mg/day) showed very low levels of both 
precursors in the trophoblast and showed an absence of phenylalanine in the inner 
cell mass. A striking observation is the fact that these were the only blastocysts 
in the entire experiment which showed a total or complete absence of uridine in 
the inner cell mass. The same pattern of uridine distribution was observed with 
both concentrations of uridine tested. Cyclohexamide (5 figlml and 50 figlml caus- 
ed a 40% and 100% inhibition, respectively, of leucine incorporation. Actinomycin 
D (0.01 figlml and 0.1 figlml) caused 44% and 88% inhibition, respectively, of 
3 H-uridine incorporation and a marked depression of 3 H-phenylalanine into pro- 
tein after three hours of incubation. 

Discussion 

Earlier studies (12) in our laboratory demonstrated that thyroxine in dosages 
as low as 8 /xg/day was able to counteract the detrimental effects of progesterone 
deficiency upon survival of blastocysts during experimentally induced delay of 
implantation. On the other hand, thyroidectomy worsened the effects of pro- 
gesterone deficiency. Although it is known that the diapausing blastocyst shows 
decreased metabolic and synthetic activities (15) our earlier work demonstrated 
that the thyroid effect upon the blastocyst is exerted during the progesterone- 



Table 2. Summary of Autoradiographs Showing In Vitro "Incorporation" of 

Radioactive Precursors into Experimentally Delayed Blastocysts from 

Progesterone-Maintained Rats with Altered Thyroid Activity 



Relative Concentration of Precursor 



Experiment*! Group 



Trophoblast 


Inner Cell Mass 


Phenylalanine 


Uridine 


Phenylalanine 


Uridine 


+++ 


++ 


- 


+/- 


+++ 


+++ 


- 


+/- 


++ 


+ 


+++ 


+++ 


+ 


+++ 


+/- 


+++ 


+ 


+ 


+ + 


++ 



Euthyroid 2.0 mg progesterone 
Euthyroid 0.4 mg progesterone 
Hyperthyroid 2.0 mg progesterone 
Hyperthyroid 0.4 mg progesterone 
Hypothyroid 2.0 mg progesterone 
Hypothyroid 0.4 mg progesterone 



Autoradiographs were analyzed for at least five blastocysts in each experimental group. 



+++ = heavy 
++ = medium 

+ = light 
+/- - very light 
- = absent 



140 Indiana Academy of Science 

dependent delay period. Possibly this effect was exerted by way of the thyroid 
hormone influencing the uterus since thyroid hormone was shown to exert an 
effect upon uterine physiology during the delay period (11). No information had 
been obtained concerning whether the blastocysts themselves showed metabolic 
differences in response to the thyroid hormone. Thus, the current studies were 
conducted to determine whether the maternal thyroid status influences the in 
vitro synthesis of protein and RNA in non-delayed blastocysts and in delayed 
blastocysts. 

The 14 C-leucine uptake and incorporation studies with blastocysts which 
were flushed from the uteri on Day 5 show that the maternal thyroid status does 
significantly influence protein synthesis in the non-delayed blastocyst. The incor- 
poration of leucine into protein has been used by a number of investigators (e.g. 
14, 4, and 17) as a useful means of assessing protein synthesis in preimplantation 
mammalian embryos. Perhaps in these intact (non-delayed) rats this effect of 
thyroid hormone on protein synthesis in the blastocyst may help explain the find- 
ings of many investigators who have shown hypothyroidism to have detrimental 
effects upon litter size while thyroid hormone administration was shown to have 
beneficial effects as reviewed by Leatham (14). 

The leucine incorporation and uptake studies were conducted utilizing 
blastocysts from intact (non-delayed) rats since large numbers of blastocysts were 
desirable for the protein synthesis studies (experimental delay of implanatation 
leads to a 50% decrease in the number of available blastocysts), and since the 
diapausing blastocyst has been shown (15) to reduce its metabolic activities. These 
investigators and Britton-Casimuri et al. (5) demonstrated that diapausing rat 
blastocysts show increased metabolic activity after 4-6 hours in an appropriate 
culture medium (removed from uterine influences). For the autoradiographic 
studies of the incorporation of protein and RNA precursors we utilized delayed 
blastocysts which were flushed from the uteri on Day 8 and incubated in BMOC-2 
for five hours. 

This relatively short incubation time does not fully restore the blastocyst to 
metabolic levels comparable to non-delayed blastocysts. However, it does provide 
an opportunity to obtain information about blastocysts that are recovering from 
the progesterone-dependent delay period during which thyroid hormone was 
demonstrated to exert an effect in our earlier studies. Experiments using various 
recovery times will be interesting to conduct and are planned, but such 
blastocysts will be less similar to delayed blastocysts. 

Although blastocysts from euthyroid rats with progesterone deficiency showed 
no differences in the incorporation and distribution of the precursors, the hyper- 
thyroid condition in both progesterone levels did cause an increase in both precur- 
sors in the inner cell mass region. The data from the leucine uptake and incorpora- 
tion studies using non-delayed blastocysts cannot be compared closely with the 
autoradiographic studies, but the increased protein and RNA synthesis during 
hyperthyroidism in these delayed blastocysts is in line with the increased protein 
synthesis seen in the non-delayed blastocysts from hyperthyroid rats. Also in line 
with the leucine incorporation studies, the autoradiographic studies with 
blastocysts from hypothyroid rats showed the lowest levels of incorporation of the 
RNA and protein precursors. The complete absence of the radioactive amino acid 
and uridine in the trophoblast area of the blastocysts from progesterone-deficient 
hypothyroid rats may be an indication of a blastocyst condition that contributes to 
extremely low survival of embryos in rats of this treatment during delayed im- 
plantation as reported by Holland et al. (10) 



Cell Biology 141 

Recently, it was shown (8 and 9) that the preimplantation embryos of the rat 
and rabbit synthesize steroid hormones; and it has been suggested (9) that these 
steroid hormones of the blastocyst are critical for preimplantation embryogenesis. 
It also has been suggested (1) that blastocysts of the rabbit accumulate steroids 
from the uterine fluid. Some of the evidence that the blastocyst is able to syn- 
thesize progesterone and estrogen was obtained by histochemically 
demonstrating the activity of 3B hydroxysteroid dehydrogenase, a key enzyme in 
steroidogenesis. Interestingly, Dickman et al. (9) reported that in their studies 
with rabbit blastocysts, for most embryos, the 3B-HSD activity was stronger in 
the inner cell mass (embryonic disc) than in the trophoblast. In our current ex- 
periments, the blastocysts from progesterone deficient hypothyroid rats were the 
only ones which showed an absence of both protein and RNA precursor incorpora- 
tion in the inner cell mass and, in delayed implantation studies, this group is the 
one for which progesterone deficiency is most detrimental. Preliminary studies of 
3B-HSD synthesis in our blastocysts indicate the possibility that progesterone- 
deficient hypothyroid rats show a lower level of this enzyme in their blastocysts. 



Acknowledgements 

This research was supported by the National Science Foundation Grant GB-8241 and 
by a Creative and Research Fellowship from Omega Psi Phi, Inc. to J.P.H. 



Literature Cited 

1. Borland, R. M., G. F. Erickson, and T. Ducibella, (1977). Accumulation of 
steroids in rabbit preimplantation blastocysts. J. Reprod. Fert. 49, 219-224. 

2. Brinster, R. L. (1970). Mammalian embryo culture. In: The Mammalian Ovid- 
cut, pp. 419-441. Ed. E.S.E. Hafez and R. J. Blandau. The University of Chicago 
Press, Chicago, Illinois. 

3. Brinster, R. L. (1971). Uptake and incorporation of amino acids by the 
preimplantation mouse embryo. J. Repord. Fert. 27, 329-338. 

4. Brinster, R. L., J. L. Wiebold, and S. Brunner, (1976). Protein metabolism in 
preimplantation mouse ovaries. Devel. Biol. 51, 215-224. 

5. Britton-Casimiro, V., J. L. Brun, and A. Psychoyos, (1976). Uptake and incor- 
poration of [ 3 H] uridine by normal or diapausing rat blastocysts after various 
periods of culture. J. Reprod. Fert. 46, 477-448. 

6. Cochrane, R. G. and R. K. Meyer, (1957). Delayed nidation in the rat induced by 
progesterone. Proc. Soc. Exp. Biol. Med. 96, 155-159. 

7. Daentl, D. L. and C. J. Epstein, (1971). Developmental interrelationships of 
uridine uptake, nucleotide formation and incorporation into RNA by early mam- 
malian embryos. Devel. Biol. 24, 428-442. 

8. Dey, S. K. and Z. Dickman, (1974). A 5 -3B Hydroxysteroid dehydrogenase acti- 
vity in rat embryos on days 1 through 7 of pregnancy. Endocrinology 95, 
321-322. 

9. Dickman, Z., S. K. Dey, and J. S. Gupta, (1975). Steroidogenesis in rabbit 
preimplantation embryos. Proc. Natl. Acad. Sci. 72, 298-300. 

10. Holland, J. P., J. M. Dorsey, N. N. Harris, and F. L. Johnson, (1967). Effect of 
thyroid activity upon delayed implantation of blastocysts in the rat. J. Reprod. 
Fert. 14, 81-85. 

11. Holland, J. P., F. L. Calhoun, N. N. Harris, and N. W. Walton, (1968). 
Uterine alkaline phosphatase and blastocyst implantation during altered 
thyroid activity. Acta Endocr., Copenh. 59, 335-343. 

12. Holland, J. P., J. M. Finley, R. D. Kazwell, and F. L. Meshberger, (1970). 
Progesterone-dependent blastocyst survival during altered thyroid activity 
in the rat. J. Reprod. Fert. 23, 143-146. 



142 Indiana Academy of Science 

13. Leathem, J. H. (1972). Role of the thyroid. In: Reproductive Biology, pp. 
857-876. Eds. H. Balin and S. Glaser. Excerpta Medica, Amsterdam. 

14. Monesi, V. and V. Salfi, (1967). Macromolecular synthesis during early 
development in the mouse embryo. Exp. Cell Res. 46, 632-635. 

15. Rath, N. C. and A. Psychoyos, (1976). In vitro incorporation of [ 3 H]-thymi- 
dine by diapausing rat blastocysts. J. Reprod. Fert. 48, 225-227. 

16. Skalko, R. G. (1971). Methods for histologic and autoradiographic analysis of 
the early mouse embryo. In: Methods in Mammalian Embryology, p. 241. Ed. 
J. C. Daniel, Jr., W. H. Freeman Co., San Francisco, California. 

17. Tasca, R. J. and N. HlLLMAN, (1970). Effects of actinomycin D and cyclohex- 
imide on RNA and protein synthesis in cleavage stage embryos. Nature, Lon- 
don, 225, 1022-1025. 



Chemically Induced Hepatocellular Inclusions 
Associated with Tumorigenicity in the Liver of the Rat 

S. D. Barnard, S. D. Warner, W. J. Caldwell, and F. Parenti 
Dow Chemical, Indianapolis, Indiana 46268 

Phthalazino [2,3-b] phthalazine 5 (14H)-12 (7H) dione is a product of the phar- 
macologic research and development program of Gruppo Lepetit, Milano, Italy a 
subsidiary of the Dow Chemical Company, Midland, Michigan. This experimental 
drug, also known as L-5418, has demonstrated anti-inflammatory activity in 
several experimental animal models and has shown effectiveness in the treatment 
of rheumatoid arthritis in humans (8). 

Short term animal studies were conducted in the Italian facility to evaluate 
the potential for any toxicity with L-5418. Oral LD 50 values were found to be 
several grams per kilogram of body weight in the dog, monkey, mouse, rat and 
rabbit. An oral LD 50 value of 10 g/kg was determined for the Wistar rat. Results 
of subchronic oral toxicity studies of L-5418 also showed a low order of toxicity. A 
four-week test involving daily 500 mg/kg administration of L-5418 by gavage to 
Wistar rats established increased liver and adrenal weights as the only adverse 
effects. In six-month studies using Wistar and Carworth Farm rats, doses of 75 to 
600 mg/kg/day given by gavage produced statistically significant increases in liver 
weights for males at all dose levels and females at the 300 and 600 mg treatment 
levels. Also, 600 mg/kg caused a decreased body weight gain in females. Oral 
doses of 600 mg/kg/day were associated histopathologically with centrolobular 
cloudy swelling and periportal fatty change in parenchymal cells of the liver. 

Subsequently, at Dow's Human Health and Consumer Products Department 
in Indianapolis, a two-year L-5418 study was conducted using dietary levels of 75 
to 600 mg/kg/day in Sprague-Dawley rats to evaluate any possible lifetime tox- 
icologic/tumorigenic effects. An increased incidence of hepatocellular carcinoma 
was observed in females administered 150 mg/kg and greater and males given 300 
mg/kg and greater. Because of the tumorigenic effect, further development of 
L-5418 was discontinued. This report relates the morphological observations made 
of liver tissue collected during the course of this two-year experiment. 

Materials and Methods 

Young male and female Sprague-Dawley rats purchased in the weight range 
of 100-120 g were used (Harlan Industries, Cumberland, Indiana). The animals 
were housed in a single room under controlled conditions of temperature and 
humidity and were given Wayne® Lab Blox® meal (Allied Mills, Inc., Chicago, 
ID and water ad libitum. The rats were randomized into a control and four treat- 
ment groups each consisting of 70 rats/sex and then placed on test diets for- 
mulated to provide doses of 0, 75, 150, 300, and 600 mg/kg/day of L-5418. 

Rats were sacrificed while under methoxyflurane anesthesia by exsanguina- 
tion from the heart after 6, 12, 18, and 24 months of treatment. A detailed 
necropsy of each animal was done and a complete list of tissues, including liver, 
was preserved for microscopic examinations. 

Light Microscopy. Liver tissues were preserved in 10% buffered formalin fixative 
and routinely processed through paraffin embedment. Sections of liver and other 
tissues were stained with hematoxylin-eosin for microscopic examination. 

Electron Microscopy. Excised liver tissue was prepared for the preservation of 

143 



144 Indiana Academy of Science 

ultrastructural morphologies as follows: Samples were minced in osmium tetrox- 
ide buffered with barbital to pH 7.4. After 90 minutes of fixation, the specimens 
were treated en bloc with uranyl acetate, dehydrated through a graded series of 
acetones, infiltrated and embedded using Epon-Araldite (3). 

Additionally, cytochemical techniques for localizing the endoplasmic 
reticulum-marking enzyme, nucleoside diphosphatase, were applied to liver 
specimens collected at the 24-month sacrifice. Hepatic samples were fixed for a 
period of 1 hour at approximately 4°C. Following washes in cold 0.15 M cacodylate 
buffer and 7.5% sucrose, the specimens were cut into 40 \i sections using a Smith- 
Farquhar TC-2 tissue chopper. Sections were subsequently reacted for 2 hours at 
37°C in the medium of Novikoff and Goldfischer (4). After post-treatment in 
osmium tetroxide, the specimens were dehydrated in graded acetones and embed- 
ded in Spurr's epoxy (12). Controls treated in substrate-deficient reaction medium 
were also processed. 

The polymerized preparations were trimmed, then sectioned, utilizing a dia- 
mond knife-equipped Porter-Blum MT-2 ultra-microtome. Sections were collected 
on copper specimen grids, contrast enhanced with uranyl acetate and lead citrate 
solutions and examined using a Philips EM 300 electron microscope. 

Results 

Light Microscopy. Treatment related abnormalities, observed in the liver tissue 
of both male and female animals, were more pronounced in males. Light 
microscopy examination of the livers of males revealed a dose-response enlarge- 
ment of hepatocytes, principally in the periportal zone of the hepatic lobule. Some 
of the enlarged hepatocytes had foamy or fine fatty vacuolation of the cytoplasm. 
Several hepatocytes also contained acidophilic cytoplasmic inclusions which 
varied from small granules and ring shapes to large round or ovoid bodies enclos- 
ing similar but smaller bodies and/or lipid vacuoles. Liver cell inclusions were 
observed as early as 6 months in males of the 600 mg/kg dose level and after 12 
months of treatment, cytoplasmic inclusions could be found in males of all treat- 
ment groups. Inclusions were not observed in hepatocytes from female rats. 

Primary liver tumors, which were considered related to the ingestion of 
L-5418 in the diet, were characterized as either hyperplastic nodules (neoplastic 
nodules) or hepatocellular carcinomas (13). Liver tumors were observed in male 
and female rats after approximately 18 months of treatment. 

Electron Microscopy. Ultrastructural examinations of hepatocytes from L-5418 
treated rats revealed increased free ribosomes, occasional megamitochondria and 
markedly proliferated smooth endoplasmic reticulum, the latter often exhibiting 
vesicular profiles with enlarged cisternae. The most striking morphological 
change observed, however, was a frequently-occurring intracellular inclusion 
body. In profile these consisted of laminated smooth membrane bands arranged 
concentrically, generally surrounding clusters of lipid droplets (Figure 1). Addi- 
tionally, the sequestered lipid droplets were often individually circumscribed by 
smooth membrane. Examinations of the peripheries of the inclusions disclosed 
continuities with the endoplasmic reticulum of hepatocytes. At times direct 
association with rough membranes could be seen though the inclusions were in- 
variably composed of smooth membrane (Figure 2). In these instances, ribosomal 
detachment was apparent at points of transition to the smooth membrane of the 
lamellar inclusions. In section, the inclusion often measured more than 14 /* in 
diameter and the central fat cluster frequently consisted of more than 15 droplets. 



Cell Biology 



145 




C 



Figure 1. Serial sections of liver tissue from a male rat treated with 600 mg/kg/- 
day L-5418 for six months. 

A. Light microscopy of a toluidine blue-stained thick epoxy section. One of 
several hepatocellular inclusions is labelled (arrow). X 480. 

B. Low magnification electron micrograph. The same inclusion is marked 
(arrow). Other inclusion bodies are clearly distinguishable as well as large 
numbers of highly osmiophilic lipid droplets. X 880. 

C. A portion of the inclusion labelled in A and B is shown. Highly 
vesiculated SER can be seen within the inclusion. Smooth membrane 
laminated bands comprise much of the anomaly. X 13,210. 



146 



Indiana Academy of Science 




ias SaS 

FIGURE 2. Liver tissue from a male rat treated for two years with 300 mg/kg/day 
L-5418. Note the continuity of the lamellar inclusion smooth membranes with the 
paired, ribosome-studded membranes of the RER. Marked vesiculation of the 
SER can be seen. X 9710. 



Serial toluidine blue-stained epoxy sections for light microscopy and ultra-thin 
sections for electron microscopy suggested that the concentric lamellar inclusion 
is the counterpart of the acidophilic body described in hematoxylin-eosin paraffin 
sections. 

Liver tissues processed for the demonstration of nucleoside diphosphatase 
activity exhibited, as normally expected, distinct enzyme localizations within the 
nuclear envelope and endoplasmic reticulum (Figure 3). Furthermore, lamellar in- 
clusions showed a similar high degree of enzymatic activity. In properly oriented 
sections, continuity of reaction product was observed between endoplasmic 
reticulum and the inclusion body (Figure 4). 

Discussion 

Hepatocellular inclusion bodies of the type reported herein are not new find- 
ings. They have been described after treatments with a number of chemically 
dissimilar compounds (2). Stenger, in a review of hepatocyte smooth endoplasmic 
reticulum responses to drug administration, reported that many toxic substances 
can cause the development of extraordinary lamellar formations of endoplasmic 
reticulum. These structures were described as consisting of concentrically dis- 
posed, paired membranes surrounding cores often containing one or several amor- 
phous, osmiophilic lipid bodies (14). It would appear that the hepatocellular inclu- 
sion induced by L-5418 treatment is like the one reviewed by Stenger. Several 
terms have been applied to these inclusions, e.g., myelin figures, fingerprints, 
membrane arrays. For the remainder of this discussion the term concentric 
lamellar inclusion body (CLIB) shall be espoused (2). 



Cell Biology 



147 











FIGURE 3. Nucleoside diphosphatase localizations can be visualized as electron 
dense, lead phosphate reaction product within nuclear envelopes and cisternae of 
the smooth and rough endoplasmic reticulum. Microbodies, lysosomes, mitochon- 
dria, and Golgi show no reactivity. X 6700. 










FIGURE 4. Liver tissue from a male rat administered 300 mg/kg/day L-5U18 for two 
years. Two lamellar inclusion bodies are illustrated both showing rich nucleoside 
diphosphatase reaction product. Presented are suggestions of paired inclusion 
membranes and inclusion continuities with the intacellular retriculum. X 9710. 



148 Indiana Academy of Science 

Two explanations regarding the nature of CLIB's have been proposed: (A) 
the inclusion is a degenerative structure representing a form of elaborate 
autophagic vacuole; or (B) the inclusion is a regenerative alteration composed of 
functional endoplasmic reticulum. The cytochemical observations reported in the 
present study and by others suggest an endoplasmic reticulum origin for the 
CLIB membranes (14). Furthermore, published data indicate the activities of 
membrane-bound enzymes, including nucleoside diphosphatase, are sharply reduced 
in hepatocytes after treatments with compounds producing peroxidation of in- 
tracellular membranes (1, 9, 15). Peroxidation of biological membrane lipids is con- 
sidered an integral part of cell damage and of many toxic processes (11). In this 
regard, the rich nucleoside diphosphatase presence within L-5418-induced CLIB's 
probably demonstrates non-degenerate, enzymatically intact inclusion reticulum. 
These findings support the hypothesis that in response to an exogenous chemical 
compound, the cell increases production of smooth membrane and associated drug- 
metabolizing enzymes. In events resulting in CLIB's, a specialized type of en- 
doplasmic reticulum hypertrophy occurs perhaps due to a concentration of the 
chemical to be metabolized within lipid droplets. 

It has become apparent, with increased published data, that a large number 
of the chemical substances causing CLIB's have also demonstrated carcinogenicity 
in animal tests, e.g., nitrosamines, DDT, PCB's, PBB's, thioacetamide, 
N-2-fluorenylacetamide, azo compounds, aflatoxins, ethionine, etc. (6). Based upon 
the findings of this study, L-5418 can be added to the list. In regard to another 
ultrastructural anomaly, Reddy has recently reported that chemically dissimilar 
compounds which induce rat liver peroxisome proliferation also cause 
hepatocellular carcinoma, and this prompts concern over the potential car- 
cinogenicity of peroxisome proliferators as a class (10). Similar statements would 
apply to compounds inducing CLIB formation. 

CLIB's within hepatocytes from rats treated with experimental chemicals 
may occur very early in testing. For example, Herdson, et al, reported "myelin 
figure" inclusions after 7 days of dietary treatment with a thiohydantoin com- 
pound (5). Also, Koff, et al, found "laminated smooth membrane-containing 
bodies" in hepatocytes collected 24 hours after the administration of a single injec- 
tion of D-galactosamine (7). In drug research, the orderly development of drug 
candidates includes long term toxicity/oncogenicity studies. These testings are ex- 
pensive requiring a great deal of laboratory space and commitments in person- 
nel—often to the point of creating "bottlenecks" which impede the progress of 
other drug candidates. Therefore, the early detection of meaningful morphologic 
indicators, in short-term toxicity evaluations, can be very important in the drug- 
selection process. These results suggest hepatocellular concentric lamellar inclu- 
sion bodies to be significant early indicators. 



Literature Cited 

1. Acheampong-Mensah, D. and G. Feurer. 1975. Action of phenobarbital and 
carbon tetrachloride on hepatic microsomal inosine diphosphatase in the rat. 
Res. Communic. Chem. Pathol Pharmacol 11:475-485. 

2. Ghadially, F. N. 1975. Ultrastructural Pathology of the Cell. Butterworths, 
Boston, Massachusetts. 

3. Hayat, M. A. 1972. Basic Electron Microscopy Techniques. Van Nostrand 
Reinhold Co., New York, New York. 

4. Hayat, M. A. 1973. Electron Microscopy of Enzymes, Vol. 1. Van Nostrand 
Reinhold Co., New York, New York, 67 p. 



Cell Biology 149 

5. Herdson, P. B., P. J. Garvin and R. B. Jennings. 1964. Reversible biological 
and fine structural changes produced in rat liver by a thiohydantoin com- 
pound. Lab. Invest. 13:1014-1031. 

6. Hruban, Z. 1970. Ultranstructure of hepatocellular tumors. J. Toxicol 
Environ. Health 5:403-431. 

7. Koff, R. S., L. J. Davidson, G. Gordon and S. M. Sabesin. 1973. D-galactosa- 
mine hepatotoxicity III. Normo-active smooth endoplasmic reticulum and 
modification by phenobarbital. Exp. Mol. Pathol. 19:168-177. 

8. Nicolis. F.B., L. Schiavetti, F. Porzio, A. Manzini, M. Marchetti and G. 
Acocella. 1974. Therapeutic activity of diftalone in rheumatoid arthritis; 
short-term controlled evaluation. Int. J. Clin. Pharmacol. 10:239. 

9. Pasquali-Ronchetti, I., A. Bini, B. Botti, G. DeAlojsio, C. Fornieri and V. 
Vannini. 1980. Ultrastructural and biochemical changes induced by pro- 
gressive lipid peroxidation on isolated microsomes and rat liver endoplasmic 
reticulum. Lab. Invest. 42:457-468. 

10. Reddy, J. K., M. S. Rao, D. L. Azarnoff and S. Sell. 1979. Mutagenic and 
carcinogenic effects of a hypolipidemic peroxisome proliferator, 
[4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio] acetic acid (Wy-14,643), in rat and 
mouse liver. Cancer Res. 39:152-161. 

11. Sagai, M. and A. L. Tappel. 1979. Lipid peroxidation induced by some 
halomethanes as measured by in vivo pentane production in the rat. Toxicol 
Appl. Pharmacol. 49:283-291. 

12. Spurr, A. R. 1969. A low viscosity epoxy resin embedding medium for elec- 
tron microscopy. J. Ultrastruct. Res. 26:31. 

13. Squire, R. A. and M. H. Levitt. 1975. Report of a workshop on classification 
of specific hepatocellular lesions in rats. Cancer Res. 35:3214-3223. 

14. Stenger, R. J. 1970. Organelle pathology of the liver. Gastroenterology 
58:554-574. 

15. Wills, E. D. 1971. Effects of lipid peroxidation on membrane-bound enzymes 
of the endoplasmic reticulum. Biochem. J. 123:983-991. 



Evidence for a Trans-Plasma Membrane Electron Transport 
System in Plant Cells 

Theodore A. Craig and Frederick L. Crane 

Department of Biological Sciences, Purdue University, 

West Lafayette, Indiana 47907 

Introduction 

Dormandy and Zarday (1) demonstrated that extracellular reduction of ferri- 
cyanide occurred with intact erythrocytes and it was proposed by Mishra and 
Passow (2) that this reduction occurred via a transmembrane dehydrogenase, 
which transferred electrons from an internal redox donor to the external ferri- 
cyanide. Orringer and Roer (3) have proposed that ascorbic acid is a redox carrier 
across the plasma membrane of erythrocytes and is responsible for external ferri- 
cyanide reduction. The presence of NADH-ferricyanide reductase has been 
established in the plasma membranes of many types of cells (4, 5) and recently ex- 
tracellular ferricyanide (FeCN) reduction has been observed in yeast cells by 
Crane et al. (6), who suggest an electron transport system in the plasma mem- 
brane is responsible. In this paper we present evidence for extracellular FeCN 
reduction via a transmembrane electron transport system in the plasma mem- 
brane of intact carrot cells. The rate is stimulated by cations and is accompanied 
by an increased proton release from the cells. 

Materials and Methods 

A carrot cell culture line obtained from Drs. P.M. Hasegawa and R.A. 
Bressan of the Purdue Horticulture Department was grown in liquid suspension 
culture. Cells were harvested in the logarithmic phase of growth and washed with 
.10 M sucrose. The suspension subsequently was aerated and aliquots used as 
needed. Potassium ferricyanide and Na 3 V0 4 were obtained from Fisher Scientific 
Co., sodium ferricyanide from K and K division of ICN Pharmaceuticals, Inc. 
Oxone, a monopersulfate oxidant, was from Dupont. Tris and Mes buffers and 
PCMB (p-chloromercuribenzoate) and NEM (N-ethylmaleimide) were obtained 
from Sigma Chemical Co., sucrose from Mallinckrodt. An Aminco DW-2a spec- 
trophotometer was used to measure ferricyanide reduction using the dual beam 
mode with a reference at 500 nm to reduce problems with turbidity. The sample 
was stirred continuously with a magnetic stirring assembly. pH measurements 
were done using an Orion model 701 A pH meter with a Corning glass combina- 
tion electrode. A millimolar extinction coefficient of 1.0 was used for potassium 
and sodium ferricyanide. 

Results 

Experiments were done to determine whether ferricyanide (FeCN) reduction 
was due to FeCN entering into cells with subsequent reduction. Cells were 
allowed to reduce FeCN under normal assay conditions (pH 7.0 Tris-Mes-HCl 0.05 
M, 0.047 M sucrose) and total ferri-ferrocyanide content was determined in the 
supernatant after cells were removed by centrifugation. Table I shows that nearly 
all of the FeCN added to the cells was reduced after 10 minutes and that an 
average of approximately 98% of this could be accounted for in the supernatant 
and not in the cells after centrifugally removing the cells. Figure 1 shows the ef- 
fects of cations on FeCN reduction. CaCl 2 and MgCl 2 gave a maximum stimulation 

150 



Cell Biology 



151 



TABLE I. Ferricyanide Recovery in Extracellular Medium Following 
Ferricyanide Reduction by Cells 





Ferricyanide 


Ferricyanide 


Recovery 


Recovery Normalized 




Reduced After 


Recovered in 




to Sucrose 




10 minutes 


Supernatant 




Control 




(/iMoles) 


(jiMoles) 


(%) 


(%) 


Buffer, sucrose 


.207 


.195 


94.2 


98.8 


Buffer, sucrose 


.207 


.197 


96.7 


101.4 


Buffer + Cells 


.182 


.174 


95.6 


100.2 


in sucrose 










Buffer + Cells 


.191 


.178 


93.2 


97.7 


in sucrose 










Buffer + Cells 


.191 


.173 


90.6 


95.0 



in sucrose 



TABLE I. Assays contained 0.05 M Tris-Mes-HCl, pH 7.0, and 0.047 M sucrose. A small amount of sodium 
ascorbate was added to the two buffered sucrose controls to reduce the ferricyanide for subsequent deter- 
mination of recovery oberved with no cells present. 7.02 mg dry wt of cells in 1.5 ml volume were allowed to 
reduce FeCN for 10 min., after which cells were removed by a 4 min. spin in a Beckman 152 microfuge. Oxone 
oxidized all ferrocyanide and the change in optical density on ascorbate addition was used for measuring 
total FeCN. 

of the rate of 40% and 35% while KCI showed a stimulation of only 6% at the 
highest concentration examined. The ability of cells to reduce FeCN was unaf- 
fected by sulfhydryl reagents PCMB or N-ethyl maleimide. Orthovanadate, a 
plant plasma membrane ATPase inhibitor (7), had no effect on the rate of FeCN 
reduction in the concentration range 0.5-5.6 mM. 

NADH is a possible source of intracellular reducing equivalents for external 











40 




• CaCI 2 
■ MgCl 2 
A KCI 


30 






20 ■ 






10 ■ 


/f— 


_ 


01 
-in 





10 

Salt Concentration (mM) 



15 



20 



Figure 1. Salt Effects on Ferricyanide Reduction by Whole Cells. For KCI 
effects Na 3 Fe(CNI 6 (0.81mM) was added and for MgCl z and CaCl 2 K 3 Fe(CN) 6 
(0.81mM) was added to cells in 50mM Tris-Mes-HCl pH 7.0, and 0.047 M sucrose. 



152 Indiana Academy of Science 

Table II. Ethanol Stimulation of Starved Carrot Cells and Pyrazole Inhibition of 

Stimulation 

FeCN Reduced/MIN/ Effect 

MG DRY WT 

(NMOLE) (%) 

A. l,(-)Pyrazole 2.15 

+ 5X Ethanol 3.29 +54 

+ 5X Ethanol 4.03 +88 

2.(->Pyrazole 2.37 

+ 10X Ethanol 4.46 +88 

3.( + (Pyrazole (.1 M) 2.19 

+ 10X Ethanol 2.37 + 8 

4.(-)Pyrazole 2.42 

+ 10X Methanol 2.25 - 7 



+ 10X N-Propanol 2.30 + 10 

1.91 
+ 10X Ethanol 3.24 + 70 

5.( + IPyrazole (.1 M) 2.39 

+ 10X Ethanol 2.37 - 1 

B. l.(+)Pyrazole(.lM) 
+ 5X Ethanol 

2.(-)Pyrazole 
+ 5X Ethanol 

3.(-)Pyrazole 
+ 5X Ethanol 

+ 5X 

4.( + IPyrazole (.1 M) 
+ 5X Ethanol 
+ 5X Ethanol 

TABLE II. Cells were washed and incubated in 0.10 M Na£H P0 4 , pH 7.0 for various lengths of time: A. 2.5 
hrs. B. 1 and 2-1 hr. B. 3 and 4 - 5 hrs. Assay conditions A: 0.047 M Na P0 4 , pH 7.0 B: 0.037 M Na P0 4 , pH 7.0 
A and B: 50 mM Tris-Mes- HC1, pH 7.0, 0.81 mM K 3 Fe(CN) 6 . 

FeCN reduction and this was investigated by using ethanol to try to increase the 
intracellular NADH levels in starved cells through the cytosolic alcohol 
dehydrogenase. Stimulation of FeCN reduction by ethanol up to 88% is observed 
in starved cells in Table IIA, #1 and #2 and in Table IIB, #2 and #3. Ethanol 
stimulation is clearly due to alcohol dehydrogenase activity, as seen by the ability 
of pyrazole, an alcohol dehydrogenase inhibitor, to prevent stimulation (Table IIA, 
#3 and #5, Table IIB, #1 and #4). Also methanol, a non-substrate, and n-propanol, a 
poorer substrate than ethanol for alcohol dehydrogenase fail to significantly 
stimulate, while subsequent ethanol addition does stimulate (Table IIA, #4) exter- 
nal ferricyanide reduction. 

The possibility that a reducing substance comes across the membrane to 
reduce FeCN externally was tested by measuring the ability of the supernatant 
from incubated cells to reduce FeCN. Table III shows that the supernatant had 
only 8-9% of the activity of whole cells. Some of the remaining activity might be 
due to the few remaining cells in the supernatant. 

Carrot cells exhibited H + release on addition of FeCN, as measured on a pH 



3.07 




2.66 


-13 


3.99 




4.77 


+ 20 


1.86 




2.00 


+ 8 


2.44 


+ 31 


1.78 




1.51 


-15 


1.46 


-18 



Cell Biology 



153 



TABLE III. Ferricyanide Reducing Activity in the Supernatant of Washed 

Carrot Cells 



FeCN Reduced After 


Average of 


FeCN Reduced After 


% of 




10 minutes in Presence 


Whole Cell 


10 minutes by 


Whole Cell 




of Cells 


Rate 


Supernatant from a 

Suspension of Cells 

Incubated Without FeCN 


Rate 




(pMotes) 


( M Moles) 


for 10 minutes 

(jiMoles) 







.182 
.191 
.191 



.017 
.015 



9.0 
8.0 



TABLE III. 0.12 mM FeCN was added either directly to a cell suspension or to the supernatant derived 
from a cell suspension incubated 10 minutes. Assay conditions were 50 mM Tris-Mes - HC1, pH 7.0, 0.047 M 
sucrose with 7.02 mg dry wt cells per 1.5 ml cuvette volume. After incubation for 10 minutes, cells were 
removed by centrifugation and the supernatant was assayed for ability to reduce FeCN. Incubation took 
place in the presence of al 0.3 mM KC1 or b, 1.20 mM KC1. 

Table IV. Comparison of Ferricyanide Reduction and H + Release on 
Ferricyanide Addition to Carrot Cells 





nmoles H + /min/mg dry wt 
Rate 


Net Increase 


nmoles/min/mg dry wt 


Rate 


Rate of 


Before FeCN 


After FeCN 


in Rate of 


FeCN 


Addition 


Addition 


H + Release 
on FeCN 
Addition 


Reduction 


23.3 


31.6 


8.3 


1.8 


31.1 


39.6 


8.5 


2.5 



TABLE IV. Tris-Mes-HCl, 1.0 mM, KC1 20 mM, K 3 Fe(CN) 6 0.81 mM. The pH was 7.3 at the start and 6.5 at 
the end of a run. FeCN was added to the pH or spectrophotometer cuvette 2.5 min. after cells were added to 
the buffer in the pH chamber (from which an aliquot of 1.5 ml was removed to measure FeCN reduction). 



meter at the same time that FeCN was reduced, as shown in Table IV. Cells 
exhibited a rate of H + release without FeCN which was stimulated on addition of 
FeCN. 

Table V shows that 46% of the FeCN rate was dependent on 2 presence. 
Addition to 2 to deoxygenated cuvettes clearly shows that 2 is required for max- 
imum activity. 

Discussion and Conclusions 

The evidence which has been presented suggests that FeCN is not taken up 
by cells and lack of ability to bring about FeCN reduction in cell suspension super- 
natant (Table III) suggests, although it does not unequivocally demonstrate, that 
it is not the release of reducing compounds from the cell which is responsible for 
the observed rate of FeCN reduction. The lack of inhibition by sulfhydryl 
reagents PCMB and NEM indicates that FeCN reduction is not due to oxidation of 
the sulfhydryl group of a transmembrane carrier sulfhydryl compound crossing 
the membrane, and that it is not non-specific oxidation of membrane protein 
sulfhydryl groups. 

The presence of a transmembrane electron transport system is an explana- 
tion for the FeCN reduction observed with whole carrot cells. Orringer and Roer 
(3) have argued that external FeCN reduction with human erythrocytes occurs as 



154 Indiana Academy of Science 

Table V. O2 Involvement in Ferricyanide Reduction in Carrot Cells 

nmoles FeCN Reduced/ 

min/mg dry wt Average %Change 



K 3 Fe(CN) 6 (.404mM) 




air 


3.50 


air 


3.43 


+ aeration with 2 -20 sec. 


3.84 


argon 


2.02 


+ aeration with 02-20 sec. 


3.33 


argon 


1.75 



aeration with 2 -20 sec. 3.64 



air 


3.47 


argon 


1.89 


argon 


3.49 


+ 2 





45.5 



TABLE V. Cells in 50 mM Tris-Mes-HCl, pH 7.0, and 0.047 M sucrose were either aerated or had argon 
bubbled through them for 2.5 min and then were covered, opened momentarily for FeCN addition. In certain 
cases pure 2 was bubbled through for 20 sec. 



a result of NADH oxidation within the cell. Ethanol stimulation using alcohol 
dehydrogenase reported here suggests NADH is involved in supplying reducing 
equivalents on the internal face of the plasma membrane for FeCN reduction out- 
side of carrot cells. 

The findings of Mishra and Passow (2) and Dormandy and Zarday (1) support 
the concept of a protonophoric transmembrane NADH dehydrogenase in 
erythrocytes. They showed that reduction of FeCN by intact erythrocytes was 
accompanied by proton transfer across the membrane. Here proton release 
stimulated by FeCN may be due to proton translocation across the membrane as a 
result of electron flow from a compound, such as NADH, to FeCN outside the cell. 
The non-stoichiometric nature of the proton release might be due to release from 
control of an electron transfer pathway pumping protons with another acceptor, 
possibly 2 , as the natural acceptor. Another possibility might be an effect on the 
plasma membrane H + -ATPase, stimulating it to pump protons external to the 
cell. Crane et al. (6) observed proton release, stoichiometric to external FeCN 
reduction in yeast cells, but added KCN to inhibit proton movement due to the 
plasma membrane ATPase before FeCN was added, which was not done here. 

The data showing 2 involvement in at least a part of the rate of reduction of 
FeCN is evidence that 2 could be a natural electron acceptor. Superoxide could 
be the immediate product of such an electron transfer system and this would be 
able to reduce FeCN. Such a system could be envisioned to result in superoxide 
dismutation to H 2 2 , which could be used in lignin biosynthesis. 

The stimulations by Ca + 2 and Mg + 2 are very similar both in extent and 
saturating concentrations to those seen by Crane et al. (6) with yeast external 
FeCN reduction. They saw approximately 40% stimulation, saturating at about 5 
mM, while we see here about 40% stimulation at 10 mM and about 30% stimula- 
tion at about 2.5 mM for CaC^ and MgC^, respectively. The effect of Ca + 2 could 
be explained as an increase in H + movement, resulting in an increased electron 
transport rate. Ca + 2 uptake has been shown to be coupled to proton transport in 
yeast (8, 9). 

There are evident similarities between this electron transport system in the 
carrot cell plasma membrane and the ferricyanide reduction system in yeast 
plasma membrane and the transmembrane NADH-ferricyanide reductase of 
erythrocytes. In each FeCN reduction occurs and is accompained by the transport 



Cell Biology 155 

of protons. In each as well the rate of FeCN reduction is stimulated by calcium 
and magnesium ions. There may be very real similarities in the organization of the 
electron transport systems in the plasma membranes of these three types of cells. 
Such a system may be present in the plasma membranes of a diversity of 
organisms. 

Since the NADH dehydrogenase in erythrocytes and the other animal cell 
membranes is responsive to hormones and it has been proposed that this 
dehydrogenase is involved in control of cell function (10, 11), this system may be 
affected by plant hormones and could be a key mediator for one or more plant hor- 
mones. Since the FeCN reduction rates are expressed on a per mg dry weight 
basis, NADH-FeCN reductase activity per mg plasma membrane protein would be 
considerably higher and more in line with values seen for NADH-FeCN reductase 
in isolated mammalian membranes. 

Acknowledgements 

This study was supported by N.I.H. Grant #AM25235. 

Literature Cited 

1. Dormandy, T. L. and Z. Zarday. 1965. The mechanism of insulin action: the 
immediate electrochemical effects of insulin on red-cell systems. J. Physiol. 
180:684-707. 

2. MlSHA, R. K. and H. Passow. 1969. Induction of intracellular ATP synthesis 
by extracellular ferricyanide in human red blood cells. J. Membrane Biol. 
1:214-224. 

3. Orringer, E. P. and M. E. S. Roer. 1979. An ascorbate-mediated transmem- 
brane-reducing system of the human erythrocyte. J. Clin. Invest. 63:53-58. 

4. Crane, F. L., H. Goldenberg, D. J. Morre and H. LOw. Dehydrogenases of 
the Plasma Membrane, in Subcellular Biochemistry, ed. D. B. Roodyn, Vol. 6, 
1979. Plenum, New York, pp. 345-399. 

5. LOw, H. and F. L. Crane. 1978. Redox function in plasma membranes. 
Biochim. Biophys. Acta 515:141-161. 

6. Crane, F. L., H. Roberts, and A. W. Linnane. Transmembrane ferricyanide 
reduction by yeast cells, Saccharomyces cerevisae. in preparation. 

7. Cocucci, M., A. Ballerin-Denti, and M. T. Marre. Effects of orthovanadate 
on H + and K + transport, transmembrane potential and plasmalemma AT- 
Pase activity in plant tissues, in Plant Membrane Transport: Current Con- 
ceptual Issues, eds. R. M. Spanswick, W. J. Lucas, and J. Dainty, 1980, 
Elsevier/North Holland Biomedical Press, pp 505-506. 

8. Hinnen, R. and E. Racker. 1979. Ca + 2 translocation in Ehrlich ascites tumor 
cells. Experientia 35:935. 

9. Stroobant, P. and G. A. Scarborough. 1979. Active transport of calcium in 
Neurospora plasma membrane vesicles. Proc. Nat. Acad. Sci. U.S.A. 
76:3102-3106. 

10. De Pierre, J. W. and L. Ernster. 1977. Enzyme topology of intracellular 
membranes. Ann. Rev. Biochem. 46: 201-262. 

11. Kant, J. A. and T. L. Steck. 1972. Cation-impermeable inside-out and right- 
side-out vesicles from human erythrocyte membranes. Nature NB 240:26-27. 



Teratogenic Potential of Phenytoin in Different 
Strains of Mice 

D. K. Hansen and M. E. Hodes 

Department of Medical Genetics 

Indiana University School of Medicine 

Indianapolis, Indiana 46223 

Introduction 

Phenytoin (PHT) is an anticonvulsant drug which is commonly used in the 
treatment of epilepsy. Evidence was first introduced in 1967 which suggested that 
PHT is teratogenic in humans (15). Subsequent studies demonstrated that women 
with epilepsy who had taken anticonvulsant drugs during their pregnancies had 
more frequent malformations in their offspring than either women with epilepsy 
who had not taken drugs or non-epileptic women (9). Hanson and Smith (6) describ- 
ed the fetal hydantoin syndrome, a pattern of malformation in children whose 
mothers had taken PHT during their pregnancies. It has been estimated that 11% 
of the children exposed to PHT in utero have the fetal hydantoin syndrome and an 
additional 31% have some of the features of the syndrome (7). A genetic 
predisposition for such drug-induced malformations had been suggested by 
several authors (3,7,9). However, evidence on this point is unclear. 

Several studies (2,4,8,14,22) have shown that PHT causes an increase in the 
incidence of cleft palate in various strains of mice. Gibson and Becker (5) 
speculated that differences in the genetic constitutions of the two strains that 
they examined could account for the differences in susceptibility to teratogenesis 
that they observed. The current study was undertaken to examine the possible 
role of such genetic factors. 

Materials and Methods 

A mouse colony was established from each of the following strains: A/J, 
AKR/J, BALB/c, C57BL/6, DBA/2J, and ICR. The animals were housed in plastic 
cages on hardwood bedding and were allowed food and tap water ad libitum. 
Humidity, temperature, and light: dark cycle were held constant throughout all 
experiments. Virgin females 7 to 12 weeks of age were housed with males (one 
male to two or three females). Copulation was ascertained by the presence of a 
vaginal plug which determined day of gestation. Pregnant females were removed 
from the cage and were housed together. PHT dissolved in 0.05% sodium hydrox- 
ide at a concentration of 10 mg/ml was injected intraperitoneal^ on gestation 
days 10, 11, and 12. Control animals received the vehicle by intraperitoneal injec- 
tion. In the initial experiment the dosage of PHT was different, and the drug was 
administered at 50, 75, or 100 mg/kg body weight. On day 18 of pregnancy, the 
pregnant females were sacrified by cervical dislocation, the uterine horns were 
exteriorized, and the fetuses were removed. Each fetus was weighed and examin- 
ed for gross abnormalities. One-third of all fetuses were fixed in 95% alcohol prior 
to clearing in 1% potassium hydroxide and staining in alizarin red S (Eastman 
Organic Chemicals) (24). These animals were later measured for crown-rump- 
length and transumbilical distance. The remaining fetuses were fixed in Bouin's 
fluid, sectioned and examined for soft tissue abnormalities. 

Differences between drug treated and control animals within a strain in 
number of resorptions, stillbirths, and abnormal fetuses (total includes all animals 

156 



Cell Biology 157 

with cleft lip with/without cleft palate) were tested for significance by X 1 test. Dif 
ferences for continuous variables such as body weight, length, and transumbilical 
distance were examined by Student's t test (21). The five percent level of 
significance was used in all studies. 

Results 

In order to determine if there is a dosage effect of PHT on malformation fre- 
quency, the drug was administered to pregnant ICR mice at 50, 75, or 100 mg/kg 
body weight. The highest dose was toxic to the mothers and was not further 
analyzed. PHT did not significantly affect the frequency of resorptions or stillbir- 
ths at either 50 CY 2 = 1.87) or 75 (X 2 = 1.98) mg/kg. The drug did not cause a 
significant difference in the frequency of total malformations LY 2 = 0.26) or 
orofacial anomalies [X 2 = 0) at 50 mg/kg. However, at the higher dose, there was 
a significant increase in total malformations (X 2 = 14.76, P <.01). This increase 
can be accounted for by the significant increase in orofacial anomalies U 2 = 33.24, 
P <0.01). Therefore, 75 mg/kg was the dose chosen for all subsequent ex- 
periments. 

Tables 1 and 2 show the results of the strain susceptibility experiments. The 
frequency of resorptions was most affected by the drug; four of the six strains ex- 
amined demonstrated significant increases in this variable following drug treat- 
ment. Frequency of stillbirths and transumbilical distance were the variables 
least affected by drug treatment. 



Table 1. Percentage of fetal abnormalities in 6 strains of mice after 
administration of phenytoin (75 mg/kg) to the mothers. Resorptions and still- 
births are expressed as percentage of total number of implants; total 
malformations and orofacial anomalies are expressed as percentage of number of 
live fetuses. Numbers in parentheses are total number of implants in that treat- 
ment group. Twenty pregnant females were utilized in each treatment group. 





Resorptions 


Stillbirths 


Total 
Malformations 


Orofacial 
Anomalies 


A/J 

Control (177) 
Treated (162) 


13.30 
23.81* 


2.45 
2.08 


15.34 
30.21* 


6.13 
21.88** 


AKR/J 

Control (197) 
Treated (218) 


13.20 
21.56* 


2.29 
2.29 


4.57 
1.71 


1.04 
0.47 


BALB/c 

Control (208) 
Treated (194) 


18.37 
21.09 


2.50 
1.72 


1.67 
3.45 


0.00 
0.00 


C57BL/6 

Control (160) 
Treated (162) 


7.50 
8.02 


1.33 
3.87 


5.33 
.3.87 


0.63 
0.00 


DBA/2J 

Control (160) 
Treated (148) 


7.50 
25.00** 


6.33 
3.48 


6.33 
11.30 


0.67 
1.39 


ICR 

Control (216) 
Treated (231) 


4.62 
9.52* 


1.44 
0.48 


4.31 
6.67 


0.00 
3.04* 


•Signficant at 1 


).05 level 








••Significant at 


0.01 level 









158 



Indiana Academy of Science 



Table 2. Changes in selected measurements in the fetuses of 6 strains of mice 
after administration of phenytoin (75mg/kg) to the mothers. 



STRAIN 




WEIGHT(g) 
(Mean ± SD| 


CROWN RUMPImml 
(Mean ± SD) 


TRANSUMBILICALImm) 
(Mean ± SD| 


AJ 
















Control 


0.99 ± 0.16 


20.9 


± 


9.1 


5.5 ± 2.4 




Treated 


0.83 ± 0.07** 


15.5 


± 


11.4 


4.2 ± 3.1 


AKR/J 
















Control 


1.10 ± 0.21 


22.3 


± 


8.2 


5.7 ± 2.0 




Treated 


0.99 ± 0.13* 


21.9 


± 


8.0 


5.6 ± 2.0 


BALB/C 
















Control 


1.15 ± 0.16 


24.7 


± 


1.7 


6.5 ± 0.7 




Treated 


1.01 ± 0.22* 


22.7 


± 


2.0* 


6.3 ± 0.6 


C57BL/6 
















Control 


1.04 ± 0.19 


21.4 


± 


5.6 


5.4 ± 1.9 




Treated 


0.98 ± 0.19 


22.1 


± 


5.8 


5.7 ± 1.5 


DBA/2J 
















Control 


0.90 ± 0.08 


16.8 


± 


10.2 


4.3 ± 2.6 




Treated 


1.00 ± 0.20 


20.9 


± 


7.4 


5.4 ± 1.9 


ICR 
















Control 


1.19 ± 0.22 


26.3 


± 


2.0 


6.7 ± 0.3 




Treated 


1.16 ± 0.14 


27.0 


± 


2.1 


6.7 ± 0.4 



♦Significant at 0.05 level 
•♦Significant at 0.01 level 

A/J mice were the strain most susceptible to the effects of the drug. This 
strain demonstrated signficiant increases in the frequencies of resorptions, total 
malformations, and orofacial anomalies and a significant decrease in fetal weight 
following drug treatment. C57BL/6 mice were the least susceptible and did not 
demonstrate signficiant differences between control and treated animals for any 
of the variables tested. 

Discussion 

The studies of PHT-induced teratogenesis in mice that have been done in the 
past have varied in their choice of strain as well as in other aspects of 
methodology. Particularly bothersome are the different methods of administra- 
tion of the drug to the pregnant mice. The plasma concentration of the drug is 
related to its method of administration in humans (16), and there is no reason to 
suspect that this is not also true for mice. Since this variable has been different in 
most of the studies, it is extremely difficult to compare their conclusions. In order 
to reach valid conclusions on the effects of strain and thus of inheritance, it is 
necessary to examine teratogenicity in several strains under constant conditions. 
With this in mind, we tested the teratogenic potential of PHT in six different 
strains of mice under very carefully controlled conditions. We found that of the 6 
strains we examined, the A/J was the most susceptible to the effects of PHT and 
the C57BL/6 was the most resistant. Johnston et al. (10) examined only these two 
strains and found the same order of susceptibility. We currently are testing the 
teratogenicitv of the drug in F l hybrids resulting from reciprocal crosses involv- 
ing A/J and C57BL/6 mice. 



Cell Biology 159 

The mechanism of PHT teratogenesis is unknown. The drug can be 
metabolized by the liver microsomal cytochrome P-450 monooxygenase system to 
a reactive epoxide (1) which is capable of binding to tissue macromolecules (13). 
Activation of other polycyclic aromatic hydrocarbons by metabolism via this 
pathway can lead to an increase in teratogenesis (19, 20), mutagenesis (12), 
carcinogenesis (11), and toxicity (17, 18, 23). At present, the role of genetic factors 
in these aspects of metabolism and teratogenesis is unclear. 



Literature Cited 

1. Chang, T., A. Savory and A. J. Glazko. 1970. A new metabolite of 5,5-diphenyl- 
hydantoin (Dilantin). Biochem. Biophys. Res. Commun. 55:444-449. 

2. Elshove, J. 1969. Cleft palate in the offspring of female mice treated with 
phenytoin. Lancet 2:1074. 

3. FEDRICK, J. 1973. Epilepsy and pregnancy: A report from the Oxford record 
linkage study. Brit. Med. J. 2:442-448. 

4. Fritz, H., D. Muller and R. Hess. 1976. Comparative study of the teratogeni- 
city of phenobarbitone, diphenylhydantoin, and carbamazepine in mice. Tox- 
icology 6:322-330. 

5. Gibson, J. E. and B. A. Becker. 1968. Teratogenic effects of diphenylhydantoin 
in Swiss-Webster and A/J mice. Proc. Soc. Expt. Biol, and Med. 128:905-909. 

6. Hanson, J. W. and D. W. Smith. 1975. The fetal hydantoin syndrome. J. Pediatr. 
£7:285-290. 

7. Hanson, J. W., N. C. Myrianthopoulos, M. A. S., Harvey and D. W. Smith. 
1976. Risks to the offspring and women treated with hydantoin anticonvulsants, 
with emphasis on the fetal hydantoin syndrome. J. Pediatr. #9:662-668. 

8. Harbinson, R. D. and B. A. Becker. 1974. Comparative embryotoxicity of 
diphenylhydantoin and some of its metabolities in mice. Teratology 10:237-242. 

9. Janz, D. 1975. The teratogenic risk of antiepileptic drugs. Epilepsia .7 6:159-169. 

10. Johnston, M. C, K. K. Sulk and K. H. Dudley. 1979. Genetic and metabolic 
studies of the differential sensitivity of A/J and C57BL/6J mice to phenytoin 
("Dilantin") -induced cleft lip. Teratology 19:33A. 

11. Kouri, R. E. and D. W. Nebert. 1977. Genetic regulation of susceptibility to 
polycyclic-hydrocarbon-induced tumors in the mouse, in Origins of Human 
Cancer. Hiatt, H. H., J. D. Watson and J. A. Winsten, eds. Cold Spring Harbor 
Laboratory. New York, pp.811-835. 

12. Lambert, G. H. and D. W. Nerbert. 1977. Genetically mediated induction of 
drug-metabolizing enzymes associated with congenital defects in the mouse. 
Teratology 10:147-154. 

13. Martz, F., C. Fainlinger III and D. A. Blake. 1977. Phenytoin teratogenesis: 
Correlation between embryopathic effect and covalent binding of putative 
arene oxide metabolite in gestational tissue. J. Pharmacol. Exp. Ther. 
203:231-239. 

14. Massey, K. M. 1966. Teratogenic effects of diphenylhydantin sodium. J. Oral 
Ther. Pharmacol. 2:380-385. 

15. Melchoir, J. Ci O. Svensmark and D. Trolle. 1967. Placental transfer of 
phenobarbitone in epileptic women and elimination in newborns. Lancet 
2:860-861. 

16. Penry, J. K. and M. E. Newmark. 1979. The use of antiepileptic drugs. Ann. Int. 
Med. 90:207-218. 

17. Robinson, J. R., J. S. Felton, R. C. Levitt, S. S. Thorgeirsson and D. W. 
Nebert. 1975. Relationship between "aromatic hydrocarbon responsiveness" 
and the survival times in mice treated with various drugs and environmental 
compounds. Mol. Pharmacol. 77:850-865. 

18. Shichi, H., D.E. Gaasterland, N.M. Jensen and D.W. Nebert. 1978. A h locus: 



160 Indiana Academy of Science 

Genetic differences in susceptibility to cataracts induced by acetaminophen. 
Science 200:539-541. 

19. Shum, S., G. H. Lambert and D. W. Nebert. 1977. The murine Ah locus and 
dysmorphogenesis. Pediatr. Res. 11:529. 

20. Shum, S., N. M. Jensen and D. W. Nebert. 1979. The murine A h locus: In utero 
toxicity and teratogenesis associated with genetic differences in benzo[a]- 
pyrene metabolism. Teratology 20:365-376. 

21. Steel, R. G. D. and J. H. Torrie. 1960. Principles and procedures of statistics. 
McGraw-Hill Book Company, Inc. New York. 

22. Sullivan, F. M. and P. R. McElhatton. 1975. Teratogenic activity of the anti- 
epileptic drugs phenobarbital, phenytoin, and primidone in mice. Toxicol. Appl. 
Pharmacol. «%:271-282. 

23. Thorgeirsson, S. S. and D. W. Nebert. 1977. The Ah locus and the metabolism 
of chemical carcinogens and other foreign compounds. Adv. Cancer Res. 
25:149-193. 

24. Wilson, J. G. 1975. Methods for administering agents and detecting malforma- 
tions in experimental animals, in Teratology: Principles and Techniques. 
Wilson, J. G. and J. Warkany, eds. University of Chicago Press. Chicago, 
Illinois, pp. 262-277. 



Comparison of Metastatic and Non-metastatic Transplantable 
Tumors from the Jaw Region of the Rat 

Gary R. Matyas and Dorothy A. Werderitsh, Department of Biological 

Sciences, Dorothy M. Morre, Department of Foods and Nutrition, and D. James 

Morre, Department of Biological Sciences, Department of Medicinal Chemistry 

and Pharmacognosy and the Purdue Cancer Center Purdue University, West 

Lafayette, Indiana 47907 

Introduction 

The ability of a primary tumor to metastasize is frequently critical to the 
clinical expression of malignancy. In order for a tumor to metastasize, cells must 
detach from the primary tumor, be transported via the blood or lymphatic 
systems, and must reattach in another organ. Poste and Nicolson (15) have reviewed 
evidence that the detachment-reattachment process is due to changes in the cell 
surface. 

In an effort to clarify these changes, we have initiated studies in which 
metastatic and non-metastatic transplantable tumor lines of similar origins are 
compared. In this report, two tumors from the jaw region of the rat, one non- 
metastatic and the other metastatic, are partially characterized in terms of light 
and electron microscopy and gross morphological characteristics, as well as 
ganglioside and neutral glycosphingolipid composition. While the metastatic 
tumor line has increased levels of total gangliosides, there is a marked decrease in 
the more complex gangliosides as compared to the non-metastatic line. 

Materials and Methods 

The tumor designated JT 2 TS f was isolated initially from soft tissues in the 
area of the articulation of the maxilla and mandible of a male inbred rat of the 
CDF strain (Harlan Industries, Indianapolis) which had been fed a diet containing 
the liver carcinogen 2-acetylaminofluorene as described (12). The second tumor, 
WJT, believed to be of spontaneous origin, was isolated initially from the same 
anatomical region of a 1.5 year old male albino rat of the same strain fed a stand- 
ard laboratory diet. The tumor lines were maintained by subcutaneous implana- 
tion of approximately 10 6 tumor cells into syngeneic recipients (7). 

At the time of sacrifice, portions of each tumor were fixed in 10% buffered 
formalin, embedded in paraplast and stained with hemotoxylin and eosin for light 
microscopy. For electron microscopy, portions were fixed overnight in cold 1% 
O s 4 in 0.05 M phosphate buffer, pH 7.2, rinsed in the same buffer, dehydrated in 
a graded series of acetone, embedded in Epon, sectioned and examined and 
photographed with a Philips EM/200. 

For biochemical analyses, the tumors were minced and homogenized in four 
volumes of distilled water with a Polytron tissue homogenizer (Kinematica, 
Lucerne, Switzerland). An aliquot of the homogenate was used for protein deter- 
mination according to the procedure of Lowry (10) with bovine serum albumin as 
standard. Gangliosides and neutral glycosphingolipids were isolated according to 
a modification of the method of Ledeen (9). The homogenates were extracted with 
ten volumes of freshly distilled chloroform-methanol (1:1, v/v) overnight at 4°C 
with stirring. The extract was filtered over glass and the residue re-extracted 
with ten volumes of chloroform-methanol (2:1, v/v) overnight at 4°C. The resulting 

161 



162 Indiana Academy of Science 

extract was filtered over glass and the combined filtrates were dried with a 
rotary evaporator. The dried sample was solubilized in 100 ml of solvent A 
(chloroform-methanol-water, 30:60:8, v/v) and applied to a DEAE-Sephadex A-25 
column (acetate form) (9). After washing with solvent A, gangliosides were eluted 
with solvent B (chloroform-methanol-0.8 M sodium acetate, 30:60:8, v/v). The 
eluate was evaporated, saponified, dialyzed, lyophylized, solubilized in 10 ml 
distilled methanol and filtered over glass. Forty ml of chloroform was added to 
the final filtrate to adjust the solvent to a 4:1 ratio of chloroform to methanol. This 
mixture was applied to a Unisil column (Clarkson) and the purified gangliosides 
were eluted as described (9). Total ganglioside sialic acid was measured according 
to Warren (19) and 50 nmoles of ganglioside sialic acid of each sample were applied 
to heat activated silica gel 60 F-254 thin layer chromatography plates (E. Merck, 
Darmstadt, Germany). The plates were developed twice in chloroform-methanol- 
ammonium hydroxide-water (60:35:7.3, v/v). To allow for even passage of solvent, 
spotted plates were pre-run through the sample zone several times before the 
beginning of the separation. Visualization was with resorcinol-HCl (17). Brain 
gangliosides served as standards. For quantitation, plates were scanned with a 
densitometer (Photovolt). 

Netural glycosphingolipids were purified from the solvent A fractions of the 
DEAE-Sephadex columns. The solvent was evaporated and the residue 
resuspended in 5 ml of chloroform and applied to a Unisil column. The column was 
washed with chloroform and ethyl acetate to remove neutral lipids and interfering 
pigments, respectively. Crude neutral glycosphingolipids were eluted with 
acetone-methanol (9:1, v/v) and dried. Following mild alkaline methanolysis and 
the addition of chloroform, the samples were washed twice with methanol-water 
(1:1, v/v) and once with methanol-9% potassium chloride (1:1, v/v). The lower phase 
was dissolved in chloroform and half the sample was spotted on a thin layer 
chromatography plate as for gangliosides. The plates were developed in 
chloroform-methanol-water (70:22:3, v/v) and neutral glycosphingolipids visualized 
by spraying the plates with 50% sulfuric acid and heating (18). 

Results 

Primary tumors of either the non-metastatic or metastatic lines were white 
in color, ovoid in shape and well circumscribed (Figure 1). The non-metastatic 
JT 2 TS f tumor illustrated is representative of those used in the study and was 
removed after approximately 1 month of growth at a rate of 0.17 cm/day (Table 1). 
The representative metastatic primary tumor, WJT, was removed after approx- 
imately 3 months. 

Lung specimens taken from the tumor-bearing rats revealed metastatic foci 
only with the WJT line (Figure 2). In 47 transfers, the metastatic incidence of the 
WJT line has been a remarkable 100%. It has metastasized to the kidneys, 
diaphragm, lungs, thymus, lymphatics and the wall of the pleural cavity. In con- 
trast, the JT 2 TS f line has not metastasized in 54 transfers. 

Upon histological examination, both the metastatic WJT and the non- 
metastatic JT 2 TS f showed morphological features characteristic of squamous cell 
carcinomas (Figure 3). The principal difference was a tendency for the metastatic 
line to appear more homogeneous and less stratified than the non-metastatic line. 
Ultrastructurally, the two tumor lines were similar (Figures 4 to 6). Both had a 
poorly developed Golgi apparatus, sparse rough endoplasmic reticulum, bizarre 
mitochondrial forms, irregularly shaped nuclei and abundant lysosomes. In 



Cell Biology 




*«* 







B 



© 



FIGURE 1. Gross morphology of the primary tumors. A. Non-metastatic, fast 
growing JT 2 TSf after 1 month of growth. B. Metastatic, slow growing WJT after 
2 months of growth. Both were white in color, ovoid in shape and well circum- 
scribed. X 2. 



Table 1. 



Growth rates comparing the non-metastatic (NM), JT 2 TSf, tumor line 
and the metastatic (M), WJT, tumor line 





Growth (cm/day) 


Tumor Line 


Mean ± S.D. Range 


JT 2 TS f (NM) a 
WJT (M) b 


0.17 ± 0.04 0.11-0.22 
0.017 ± 0.005 0.01-0.025 



"Twelve passages over approximately 1 yr 
b Six passages over approximately 1.5 yr 



164 



Indiana Academy of Science 



■"PS 





Figure 2. Lung specimens from tumor bearing rats. A. Lung from JT 2 TSf. B. 
Lung from WJT. Small arrows indicate metastatic foci Large arrow denotes 
bronchus. X 2. 



Cell Biology 



165 



general, the Golgi apparatus appeared to be less well developed in the metastatic 
line whereas lysosomes appeared to be more abundant. 










XI 



1 






I- * 









• -T- 



v v 






> 



«M I 



N*v 



3i 
B 



V ■ 



& 



* 



T| 



- i &St 






: V 



• * 






Figure 3. Light micrographs demonstrating the appearance of the transplan- 
table squamous cell carcinomas. A. Non-metastatic JT 2 TSf B. Metastatic WJT. 
Generally they were similar in appearance except that cells of the WJT line 
appeared more homogeneous and less stratified than those of the JT 2 TSf line. H 
& E stain. X 500. 



166 



Indiana Academy of Science 



N 



.*' \ 




IMt 



mp-j 










/* X. 




m ■ :• - 



*$:*?* ; 



©^ 



FIGURE 4. Electron micrograph of the WJT, metastatic tumor line at low 
magnification. Note the paucity of rough endoplasmic reticulum, the presence of 
unusual mitochondrial forms (m) and numerous lysosomes (LK The Golgi 
apparatus (Gal was poorly developed, N = nucleus. Bar = 0.5ft. 




167 







.■• 







^**:i 



FIGURE 5. Electron micrograph of WJT metastatic tumor line at higher 
magnification to reveal numerous filaments in the cytoplasm (arrows). Bar = 
0.5 tL 



Total ganglioside sialic acid was approximately 3 times greater in the 
metastatic, slow-growing WJT than in the faster-growing, non-metastatic JT 2 TS f 
(Table 2). Thin layer chromatography revealed a large number of individual 
ganglioside species (Figure 8) some of which co-migrated with standards and 
others which did not. Densitometry traces of lanes corresponding to the same 
amounts of applied ganglioside sialic acid confirm what is observed visually from 
the plates in that ganglioside bands corresponding to the higher gangliosides G Dlb 
and G Tlb were much reduced or absent in extracts prepared from tumors of the 
metastatic line (Figure 9). Additionally, the metastatic line contained at least one 



Table 2. Ganglioside sialic acid and total protein comparing the non-metastatic, 
JT 2 TSf, and metastatic, WJT, tumor lines 





Wet Weight 




Protein 




Ganglioside Sialic Acid 


Tumor 


Inmoles/mg 


Designation 


Img) 


Img/g wet wt) 


(nmolesl protein) 


JT 2 TS r I a 
JT 2 TS f II a 
JT 2 TS r III a ' b (nec) 
WJT C 


27 

31 

19 

3 


2500 

3144 

520 

120 




93 
101 
27 
40 


1420 568 

1020 310 

160 320 

170 1417 



a Results from 3 different animals 

b Necrotic tumor 

c Results from pooled tumors 



168 



Indiana Academy of Science 






N 







*- 3*?*y 






^%f *$r '•-> a ■ vis* 



? ** 



'-, 



r V/' 






f L 



Tl 





AS 



Figure 6. Electron micrograph of JT 2 TSf for comparison. The nucleus (N) is 
highly lobed and irregular, lysosomes are less abundant and the Golgi apparatus 
(GaJ is better developed. Cytoplasmic filaments (arrows) are present. M = 
mitochondria. Bar = 0.5 p. 



Cell Biology 169 



B 



'M3 



Gdi< 
Gti 



JT 2 TSf JT 2 TS f Std WJT JT 2 TSf 

FIGURE 7. Thin layer chromatogram of gangliosides from rat jaw tumors. (A) 
JT 2 TF fs , (B) JT 2 TSt, (C) Standard mixed brain gangliosides, ID) WJT, (E) JT 2 TS f 
which had a high degree of necrosis. 



ganglioside band not present in the non-metastatic tumors (Figure 9). No available 
ganglioside standard was found to migrate in the region of this ganglioside band. 

Similarly increased in the metastatic line were higher neutral glycosphingo- 
lipids (Figure 10). Especially striking was the absence of neutral glycosphingoli- 
pids migrating in the vicinity of ceramide tetrahexosides (C4H). 

Discussion 

Two tumor lines, one metastatic and the other non-metastatic, of similar 
origins provide an excellent opportunity to compare various cell surface consti- 
tuents potentially related to malignancy. Also, the WJT is an unusual metastatic 
primary tumor. Generally, metastatic primary tumors are invasive, poorly cir- 
cumscribed and fast growing. The WJT does not conform to these criteria in that 
it mimics the non-invasive phenotype, is well circumscribed and slow growing. 
The many similarities of the metastatic and non-metastatic lines may eventually 
prove beneficial in understanding the metastatic process. Our observations in- 
dicate that the WJT line is able to release cells to form secondary metastases but 
still retain the gross characteristics of a benign neoplasm. 

The similarity of the two lines is apparent at both the light and electron 
microscope levels. The more abundant lysosome population of the metastatic 
isolate may be significant as a source of hydrolytic enzymes to disrupt the in- 
tracellular matrix and thus aid the disassociation and migration of invading cells. 



170 



Indiana Academy of Science 



A.JT 2 TS f (NM) 




U+tt-L-L 



> G Dlb | 



Origin 



G-rib G D1b G DlQ G M1 G D3 

Relative Mobility 



From 



Figure 8. Densitometry scans of rat jaw tumor gangliosides. (A) Scan of the non- 
metastatic JT 2 TSf from lane B of Fig. 7 (B) Scan of the metastatic WJT tumor 
(lane D of Fig. 7). Arrows indicate the relative mobilities of the standard ganglio- 
sides indicated. Note the reductions in the regions of the scans corresponding to 
the gangliosides GD lb and GT lb . ABBREVIATIONS USED: Cer = ceramide; 
G Dla = NAN-Gal-GalNAc-(NAN)-Gal-Glc-Cer; G Dlb = Gal-GalNAc-(NAN) f Gal- 
Glc-Cer; G m = GalNAc-(NAN) r Gal-Glc-Cer; G D3 = (NAN) 2 -Gal-Glc-Cer; G M1 = 
Gal-GalNAc-(NAN)-Gal-Glc-Cer; G M2 = GalNAc-(NAN)-Gal-Glc-Cer; G M3 = 
NANGal-Glc-Cer; G T = (NAN) 2 -Gal-GalNAc-(NAN)-Gal-Glc-Cer (G Tla ), NAN-Gal- 
GalNAc-(NAN) 2 -Gal-Glc-Cer (G Tlb ), or (NAN) 3 -Gal-Glc-Cer (G T3 ); G x = G Q and 
other unidentified higher ganglioside homologs; Gal = galactose; GalNAc = 
N -acetylgalactosamine; Glc = glucose; NAN = N-acetylneuraminic acid (sialic 
acid). 



The ganglioside composition of tumors appears to be highly dependent upon 
their stage of development (12). Generally, most tumor or transformed cells 
appear to have an increased level of ganglioside-associated sialic acid (8). Yet, ex- 
cept for a few rat hepatoma lines (7, 8), no studies have compared the ganglioside 
composition of metastatic and non-metastatic tumor lines of similar origins. 

At least 15 distinct ganglioside bands were resolved in the extracts prepared 
from the non-metastatic tumor line (Figure 8). At least one band was present in 
the metastatic line that could not be detected in the non-metastatic line. Since it 
did not co-migrate with any of the standard gangliosides available, it may repre- 
sent a novel structure containing fucose (11), glucosamine (2) or some other 
unusual constituent (16). Structural studies will be necessary to confirm this sup- 
position. 

More important, however, in the context of the present study was the absence 
of certain ganglioside bands in the extracts of the metastatic line (Figure 8). 



Cell Biology 



171 



Striking in this regard were the almost complete absence of gangliosides G Dlb 
and G Tlb . Both these gangliosides have been implicated recently as potential 
receptors for fibronectins, proteins that link cells to underlying substrata and may 
be important to cell adhesion (3, 5). 

Recent evidence shows that di- and trisialogangliosides inhibit the fibronec- 
tins mediated binding of cells to collagen-coated plates (5, 6). Data from our 
laboratory (14) indicate that the gangliosides G Dla , G Dlb and G T1 all bind fibronectins. 





B 



^^^^^ ^^^^ 



CMH 




CDH 




H 



C4H 



JT 2 TS, JT 2 TS ( JT 2 TS f Std 



WJT 



Figure 9. Thin layer chromatogram of neutral glycosphingolipids from rat jaw 
tumors. (A) JT 2 TS f (B) JT 2 TS< (C) JT 2 TS f which had a high degree of necrosis ID) 
Standards (E) WJT Note the reductions in the relative amount of material 
migrating in the vicinity of ChH. Abbreviations Used: CHM = ceramide 
monohexoside; CDH = ceramide dihexoside; CTH = ceramide trihexoside; C-kH 
= ceramide tetrahexoside. 



172 Indiana Academy of Science 

Additionally, there is a strong correlation between the loss of the ability of cells to 
bind fibronectins and the ability of cells to invade and form tumor metastases (1). 
This loss of ability to bind fibronectins has little or no effect on the ability of cells to 
synthesize fibronectins (1, 4). 

Evidence presented here indicates that the metastatic tumor line, WJT, loses 
the gangliosides that migrate in the region of G T1 and G Dlb . Thus it would appear 
that loss of the complex fibronectin-receptor gangliosides may contribute to the 
metastatic potential of the WJT tumor line. 

The mechanism whereby the gangliosides are lost is unknown. One possibili- 
ty is that lysosomal enzymes may be involved in their degradation. Another is 
that their biosynthesis may be blocked. In 2-acetylaminofluorene-treated rats, 
tumors exhibit a block in the branchpoint enzyme, CMP-sialic acid: G M3 sialyl- 
transferase (13) which results in a decrease in the disialo- and trisialogangliosides 
in those tissues. 

The proof of the fibronectin receptor hypothesis of ganglioside action in 
metastasis will be aided substantially by structural characterization of the miss- 
ing gangliosides and of the fibronectin-binding ability of the two tumor lines, both 
of which are in progress. We anticipate that the continued investigation of the 
metastatic and non-metastatic squamous cell carcinomas reported here will aid in 
the eventual understanding of the metastatic process during tumor progression. 

Summary 

Two rat tumors, a spontaneous jaw tumor (WJT) which, in 47 transfers, ex- 
hibited a metastatic incidence of 100% and a jaw tumor (JT 2 TS f ) induced by the 
chemical carcinogen 2-acetylaminofluorene and which has not metastasized in 54 
generations were compared. Criteria for comparison utilized both light and elec- 
tron microscopy as well as gross morphological characteristics and 
glycosphingolipid composition. The two lines exhibited similar morphologies with 
the metastatic line being more homogeneous and less stratified. Ultrastructurally, 
more lysosomes were observed in the metastatic line. Certain di- and trisialo- 
gangliosides were reduced or absent from extracts of the metastatic line that 
were present in the non-metastatic line. The reduced or missing gangliosides are 
those thought to function in fibronectin binding and cell attachment. It is sug- 
gested that both the increased lysosome content and the reduction in putative 
fibronectin receptor gangliosides may be important to the ability of the WJT line 
to form metastases. 

Acknowledgements 

Supported in part by grants from the American Cancer Society CD-52, The 
National Institutes of Health CA 18801, and a project of the Purdue University 
Agricultural Experiment Station 8546-561264. 

Literature Cited 

1. Chen, L. B., I. Summerhayes, P. Hsieh and P. H. Gallimore. 1979. Possible 
role of fibronectin in malignancy. J. Supramol. Struct. 12:139-150. 

2. Dyatlovitskaya, E. V., A. E. Zablotskaya, Y. M. Azizov and L. D. 
Bergelson. 1980. Gangliosides of calf thymus and of normal and leukemic 
bovine lymphocytes. Eur. J. Biochem. 110: 475-483. 

3. Hahn, L. E. and K. M. Yamada. 1979. Isolation and biological characteriza- 
tion of active fragments of the adhesive glycoprotein fibronectin. Cell 18: 
1043-1051. 



Cell Biology 173 

4. Hynes, R. 0., A. T. Destree, M. E. Perkins and D. D. Wagner. 1979. Cell 
surface fibronectin and oncogenic transformation. J. Supramol. Struct. 
11:95-104. 

5. Kleinman, H. K., A. T. Hewitt, J. C. Murray, L. A. Liotta, S. I. Rennard, 
J. P. Pennypacker, E. B. McGoodwin, G. R. Martin and P. H. Fishman. 
1979. Cellular and metabolic specificity in the interaction of adhesion pro- 
teins with collagen and with cells. J. Supramol. Struct. 11:69-78. 

6. Kleinman, H. K„ G. R. Martin and P. H. Fishman. 1979. Ganglioside inhibi- 
tion of fibronectin-mediated cell adhesion to collagen. Proc. Natl. Acad. Sci. 
76:3367-3371. 

7. Kloppel, T. M. and D. J. Morre. 1980. Characteristics of transplantable 
tumors induced in the rat by N-2-fluorenylacetamide: Elevations in tissue and 
serum sialic acid. J. Nat. Cancer Inst. 64:1401-1411. 

8. Kloppel, T. M m D. J. Morre and L. B. Jacobsen. 1979. Ganglioside patterns 
of metastatic and non-metastatic transplantable hepatocellular carcinomas of 
the rat. J. Supramol. Struct. 11:485-492. 

9. Ledeen, R. W., R. K. Yu and L. F. Eng. 1973. Gangliosides of human myelin: 
sialogalactosyl ceramide (G 7 ) as a major component. J. Neurochem. 
21:829-839. 

10. Lowry, 0. H., N. J. Rosebrough, A. L. Farr and R. J. Randall. 1951. Pro- 
tein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265-275. 

11. Macher, B. A., T. Pacuszka, B. R. Mullin, C. C. Sweeley, R. O. Brady and 
P. H. Fishman. 1979. Isolation and identification of a fucose-containing 
ganglioside from bovine thyroid gland. Biochim. Biophys. Acta 588:35-43. 

12. Merritt, W. D., C. L. Richardson, T. W. Keenan and D. J. Morre. 1978. 
Gangliosides of liver tumors induced by N-2-fluorenylacetamide. I. 
Ganglioside alterations in liver tumorigenesis and normal development. J. 
Natl. Cancer Inst. 60:1313-1327. 

13. Merritt, W. D., D. J. Morre, and T. W. Keenan. 1978. Gangliosides of liver 
tumors induced by N-2-fluorenylacetamide. II. Alterations in biosynthetic en- 
zymes. J. Natl. Cancer Inst. 60:1329-1337. 

14. Morre, D. J., K. E. Creek and D. M. Morre. 1981. Gangliosides and 
Tumorigenesis. In "International Cell Biology 1980-1981", H. G. Schweiger, 
ed., Springer-Verlag, Heidelberg, pp. 800-804. 

15. Poste, G. and G. L. Nicolson. 1980. Arrest and metastasis of blood-borne 
tumor cells are modified by fusion of plasma membrane vesicles from highly 
metastatic cells. Proc. Natl. Acad. Sci. 77:399-403. 

16. Stoffyn, A., P. Stoffyn and M. C. M. Yip. 1975. Chemical structure of 
monosialoganglioside G M1 b biosynthesized in vitro. Biochim. Biophys. Acta 
409:97-103. 

17. Svennerholm, L. 1957. Quantitative estimation of sialic acids. II. A col- 
orimetric resorcinol-hydrochloric acid method. Biochim. Biophys. Acta 
24:604-611. 

18. Walter, V. P., T. M. Kloppel, I. G. Deimling and D. J. Morre. 1980. Altera- 
tions in neutral glycosphingolipids from transplantable hepatomas and in 
sera of rats bearing transplantable hepatomas. Cancer Biochem. Biophys. 
4:145-151. 

19. Warren, L. 1959. The thiobarbituric acid assay of sialic acids. J. Biol. Chem. 
234:1971-1975. 



CHEMISTRY 

Chairman: Edward Miller 
Department of Chemistry, Manchester College, North Manchester, Indiana 46962 

Chairman-Elect: Donald G. Clemens 
Department of Chemistry, Goshen College, Goshen, Indiana 46526 

ABSTRACTS 
Comparisons of the Semi-empirical Molecular Orbital Treatments MINDO/3 and 
MNDO with Some Simple Phosphines. Randall K. Atkins, Paul L. Bock, John 
A. Mosbo, and Bruce N. Storhoff, Department of Chemistry, Ball State Universi- 
ty, Muncie, Indiana 47306. The semi-empirical molecular orbital computer pro- 
grams MINDO/3 and MNDO were used to obtain heats of formation data and op- 
timized geometries for all unique conformations of fourteen phosphines: PH 2 R (R 
= H, Me, Et, i-Pr, t-Bu, Ph), PHRg (R = Me, Et, i-Pr), and PRRg' (R = R' = Me, Et; 
R = Me, R' = Et; R = Et, Ph, R' = Me). The bond lengths, bond angles, twist 
angles and heats of formation predicated by the two treatments were compared, 
and the effect of differences discussed in terms of effective phosphine sizes and con- 
formational populations. Computed phosphorus-carbon rotational barrier heights 
were compared to experimental data. 

Gas Phase Chemistry of 1,3-Dithiane. John E. Bartmess, Robert L. Hays, and 
Stephen R. Wilson, Department of Chemistry, Indiana University, Bloomington, 
Indiana 47405. In a negative ion modified (ICR) mass spectrometer, we have ex- 
amined the gas phase bimolecular chemistry of the acyl anion equivalent 
1,3-dithiane. Unlike solution where only deprotonation to give 1 occurs, in the gas 





"V 



CH„ = S 



3 



phase successive eliminations are also seen. As X" is varied, the three ionic pro- 
ducts are seen to occur or not occur consistent with their thermochemistry. Similar 
chemistry is seen for the five membered ring. The difference between gas phase 
and solution is attributed to ion-pairing of the metal ion of the base with the sulfur 
in solution, thus not allowing the base access to the hydrogen leading to elimina- 
tion. The lack of a counter ion in the gas phase lets the base access either site. 

Laboratory Interfacing with the Apple II Plus Microcomputer: A Computer Con- 
trolled Titrator with High Resolution Color Graphics Display. Stanley L. Burden, 
David Woodall, Kathleen Donica, Rick Thompson, Jon Condit, Steven Beeson, 
and Douglas Taylor, Department of Chemistry and Information Science, Taylor 
University, Upland, Indiana 46989. An Apple II Plus microcomputer has been 

174 



Chemistry 175 

interfaced to an Orion 701 pH meter and a Sargent constant rate buret and soft- 
ware has been written to configure a computer controlled titrator. The high resolu- 
tion graphics capability of the Apple is used to provide a real time, color plot of the 
titration data as it is being collected. One of the user selectable options includes 
specifying a pH or millivolt reading at which the continuous titrant delivery is 
automatically changed to a dropwise addition as the endpoint is approached. 
Milliliters of titrant used and normality of titrate are computed and displayed on 
the television screen at the completion of the titration. 

Test of Nonisothermal Apparatus Suitable for the Study of Energy and Mass 
Transport Coefficients. Marshall P. Cady, Jr., Department of Natural Sciences, 
Indiana University Southeast, New Albany, Indiana 47150. A new nonisother- 
mal apparatus capable of measuring energy and mass transport coefficients of clear 
liquid mixtures has been designed and assembled. The apparatus consists of a new- 
ly designed thermal cell and a dual Savart plate Bryngdahl interferometer. 
Preliminary tests prove that nonisothermal techniques can be used to directly 
measure system properties with a high degree of accuracy and without correc- 
tions for complex energy transport phenomena associated with wall effects. 

The quality of temperature control and measurement has been verified and 
the instrument has been tested by measurement of the temperature dependence 
of the refractive index of water. The refractive index, n, is related to the 
Bryngdahl interferometric fringe number, N, by the equation: 

-dn/dT = CN/AT 

where AT is the difference in temperature between the upper and lower cell boun- 
daries and C is an instrument constant which is evaluated from isothermal in- 
terferometric images. We find that for water at 632.8 ran and atmospheric 
pressure 

-(dn/dTUO 4 = 0.313 + 0.02806T 

where T is in °C and can vary between 23°C and 38°C. Values of (dn/dT) computed 
from this equation are 2-3% lower than the standard established by Tilton and 
Taylor. However, this deviation is comparable to the standard deviation between 
all isothermal studies on water. In the absence of corrections for wall effects, 
previous nonisothermal determinations have yielded considerably lower values. 
Bryngdahl found values from 5% higher at 15°C to 12% lower at 26°C; Olson and 
Home found a value 5% lower at 25°C. 

Thermal cell construction is based upon the cell design pioneered by 
Krishnamurti but it only weighs approximately 20 pounds. It utilizes a parallel 
plate configuration in which aluminum blocks form the upper and lower cell boun- 
daries. The blocks are separated by precision nylon spacers and the block surfaces 
have been machined flat to within 0.001 inch. Front and back boundaries consist of 
optically flat plate glass held in place by nylon frames. The temperature of the 
aluminum blocks is controlled with a combination electronic heat pump and water 
heat bath. Furthermore, the vertical temperature distribution is precisely deter- 
mined with an internal thermocouple system. Temperatures are controlled to 
within approximately 0.005°C. Distinctive cell features include a relatively heavy 
weight, large distances (approximately 2 inches) between the internal heat bath 
and the cell-liquid surface, and the absence of highly polished silver mirror boun- 
daries. These features are designed to eliminate vibrations, fluid convection, exter 
nal heat flux problems, and infrared reflection problems at boundaries. 

Ultimately, we hope to simultaneously measure thermal diffusivities diffusion 



176 Indiana Academy of Science 

coefficients, and thermal diffusion factors of associating solutes in nonassociating 
solvents with this new thermal cell — Bryngdahl interferometer arrangement. The 
effort described in this paper proves the accuracy of the instrument under steady — 
state temperature conditions. To reach our goal, we must now prove that the time 
evolution of (N,T) data can be analyzed numerically for the thermal diffusivity 
coefficient of a pure liquid. These tests will be published in a future paper. 

The Schmidt Reaction of 3a, 4,5,6, -Tetrahydrosuccinimido|3,4-b|acenaphthen-10- 
one and Its Alkylated Derivatives. E. Campaigne and R. Yodice, Department of 
Chemistry, Indiana University, Bloomington, Indiana 47405. The rearrange- 
ment of 3a,4,5,6-tetrahydrosuccinimido[3,4,-b]acenaphthen-10-one and two methy- 
lated derivatives using Schmidt conditions is described. The ratio of the major pro- 
duct, 2,3,8,9-tetrahydro-3-oxo-l/f-benz[de]isoquinoline-l,9a(7^)-dicarboximide(l), to 
the minor product, 2,3,6,7-tetrahydro-3-oxo-4J7-benz-[ij]isoquinoline-4,4a(5i/)-dicar- 
boximide, under different acidic conditions is given. The ratios of analogous pro- 
ducts from the methylated derivatives are similar under similar conditions. 
Alkylations and thionations of 1 produced a variety of heterocycles, of interest as 
potential anticonvulsants. 

The Identification of Synthetic Fibers of Forensic Interest by a Combination of 
Differential Thermal Analysis and Infrared Spectrophotometry. R. J. Coffey, D. 
J. Reuland and W. A. Trinler, Department of Chemistry, Indiana State Univer- 
sity, Terre Haute, Indiana 47809. A method of differentiating between fibers of 

forensic interest by differential thermal analysis and infrared spectrophotometry 
is described. Thermograms of the first melt were used. Sample size varied from 0.1 
to 20 milligrams. In most cases fibers of different composition or from different 
manufacturers could be identified by the morphological features of the ther- 
mograms and the melting temperatures. In some cases infrared spectra also were 
required for identification. Obtaining a thermogram of the first melt enhances the 
evidential value of the analysis by recording the effect of the previous thermal 
history of the fiber on the thermogram. In some cases this allowed for the differ- 
entiation of similar fibers from different manufacturers. Care was taken to ensure 
that the sample and the comparator were run under as similar experimental condi- 
tions as possible. Where comparisons could be made the various endotherms and 
exotherms of the first melt were consistent with those reported by other investi- 
gators. 
Energy Surfaces of Sigmatropic Shifts. Joseph J. Gajewski and Kevin E. Gilbert, 

Department of Chemistry, Indiana University, Bloomington, Indiana 47405. 

Transition state structure variation in the 2,3-sigmatropic shift has been revealed 
by secondardy deuterium kinetic isotope effects at the a and 7 carbons of various 
3° -allyl amine oxides. Both extensions of Hammond's Postulate and Thornton's 
perpendicular effect are reasonable predictors of the variation observed. Secondly, 
mathematical models for sigmatropic shift energy surfaces have been developed to 
allow correlation of rate data as a function of the stabilities of product and the 
transition states for non-concerted bond breaking and bond making. 

The Chemistry of Cyanogen Compounds Degradation. Robert H. L. Howe, West 

Lafayette, Indiana 47906. The degradation of a number of cyanogen compounds 

by respective chemical, biological and physical methods is presented. Some 
experience of the author is discussed. 

Parameterization of the Empirical Molecular Conformation Approach CAMSEQ 
for Phosphines. Mark D. McIntire, Paul L. Bock, John A. Mosbo and Bruce N. 



Chemistry 177 

Storhoff, Department of Chemistry, Ball State University, Muncie, Indiana 47306. 

The empirical computational technique CAMSEQ is based on three types of 

energy functions: (1) Lennard-Jones 6-12 potential functions represent steric, non- 
bonded interactions, (2) Coulomb's law functions approximate electrostatic inter- 
actions, and (3) truncated fourier series are used to modify the torsional barriers. 
The latter has not been previously parameterized for PC and C-C bonds in 
phosphines, a necessity for optimal use of the program. This paper described the 
process for parameterization based on the available experimental data of some sim- 
ple phosphines and substituted ethanes. Use of fully parameterized CAMSEQ with 
larger phosphines was discussed and the results compared to other computational 
techniques.. 

A Study of Adenosine Deaminase in Human Serum. Thomas W. Myers and 
Pang F. Ma, Department of Chemistry, Ball State University, Muncie, Indiana 

47306. Adenosine deaminase (adenosine aminohydrolase, EC 3.5.4.4) catalyzes 

the hydrolytic deamination of adenosine to produce inosine and ammonia. Clinical 
and experimental investigations have shown that adenosine deaminase is critical in 
developing and maintaining immunological competence. The clinical and chemical 
aspects of adenosine deaminase have become quite numerous. Because of the cur- 
rent interest, the monitoring of adenosine deaminase activity is a very important 
problem. Little information has been reported concerning the activity of the dif- 
ferent molecular weight forms of adenosine deaminase in disease states or of the 
stability of the different forms during storage. Most studies determining the acti- 
vity of the individual forms of adenosine deaminase have utilized one of the follow- 
ing techniques in order to separate the different molecular forms of the enzyme: 
sucrose density gradient sedimentation, gel electrophoresis, isoelectric focusing, or 
thin-layer chromatography. These methods all present various problems when 
used in clinical studies. The difficulty and time required to perform these separa- 
tions have made duplication of results difficult in previous studies. Gel filtration 
was used in order to provide a rapid, quantitative, and reproducible separation of 
the different forms of adenosine deaminase. The gel filtration method and the 
determination of the stability of the different molecular forms of adenosine 
deaminase during storage are presented. Potential applications toward clinical 
studies also are discussed. 

Equilibria Between Diols and the NMR Shift Reagent Eu|fod) 3 . Lois M. Ounapu, 
John A. Mosbo, Paul L. Bock and Terry L. Kruger, Department of Chemistry, 

Ball State University, Muncie, Indiana 47306. The solution equilibria between 

a difunctional molecule, S, and the lanthanide shift reagent Eu(fod) 3 , L, can include 
three product forms: LS and LS 2 where the substrate functions as a monodentate 
ligand, and LS b where a single substrate functions as a bidentate ligand. An 
iterative computer program was used to fit experimental and calculated proton 
NMR chemical shifts while optimizing the parameters of equilibrium constants and 
limiting chemical shifts. Results were compared from fits of (1) LS 2 parameters 
only, (2) LS and LS b parameters only, and (3) LS, LS 2 and LS b parameters for 
strongly chelating diols (e.g., d,f-2,4-pentanediol), the more weakly chelating 
ct's-l,3-cyclohexanediol, and the non-chelating £rcms-l,3-cyclohexandiol. The viability 
of single and double equilibrium constant fits for each type of diol were discussed. 

A Comparative Study of the Interaction Between the Conversion Factors from 
Human Tissues with the Small Forms of Adenosine Deaminase from Various 
Organisms. Shashi Puttaswamy, Pang F. Ma, Department of Chemistry, Ball 



178 Indiana Academy of Science 

State University, Muncie Indiana 47306. Two molecular forms of adenosine 

deaminase have been reported from bovine livers. These two forms were found to 
differ in their size, and they have also been found to be interconvertible. Adenosine 
deaminases extracted from human tissues exhibited similar properties as well. 
Previous studies have shown the presence of a conversion factor which formed an 
aggregate with the small form (the C-form) of the enzyme, and the resulting 
enzyme complex was identified to be the large form (the A-form) of adenosine 
deaminase. Studies have been made with rat tissues in the past, and the C-form of 
the adenosine deaminase has been found to be present. On the other hand, the large 
form of this enzyme was absent in these rat tissues which were examined. Thus, it 
has been concluded that the enzyme-form distribution in rat differs from that of 
human and bovine tissues since the latter two have both the C-form and the A-form 
present in them. In this study, the conversion factor isolated from human liver will 
be tested for its universality in transforming the small form of adenosine 
deaminase from other species into the large form of the enzyme. By so doing, we 
might be able to veryify any differences that might exist between the small form of 
the enzyme present in the different species. 

The Hydrolysis of Bovine Glucagon by a Denaturant-stable Protease. Brenda L. 
Schuffman and Eric R. Johnson, Department of Chemistry, Ball State University, 

Muncie, Indiana 47306. A denaturant-stable serine protease isolated from Pro- 

nase, a commercially available proteolytic enzyme mixture, catalyzes the 
hydrolysis of bovine glucagon in the presence of 6.0 M guanidinium chloride. With 
the use of this denaturant, an average of two peptide bonds per glucagon molecule 
were catalytically hydrolyzed by the stable protease. Utilizing dansylation techni- 
ques, one of the new N-terminal amino acids produced by protease action on 
glucagon has been identified as valine. Since bovine glucagon contains only one 
valine residue, the specificity of action of this stable protease in the presence of 
denaturant includes the Phe(22)-Val(23) peptide bond of bovine glucagon. 
Dielectric and Electronic Polarizations of Substituted Metal-Acetylacetone Com- 
plexes. Eugene Schwartz, Mark Bradley, and Timothy Neufeld, Department of 
Chemistry, DePauw University, Greencastle, Indiana, 46135. The total dielec- 
tric polarizations and the electronic polarizations of a number of substituted 
symmetrical metal-acetylacetone (2,4-pentanedione) complexes were measured in 
benzene at 25.0°C. A heterodyne beat method was used to measure the dielectric 
constants at 13 KHz for the total polarizations and a differential refractometer for 
the refractive indices for the electronic polarizations. Polarizations were calculated 
by the method of Halverstadt and Kumler. Metals used were aluminum, beryllium, 
chromium (III), and iron (III). The ligands used included those with hydrogen 
(chromium only), phenyl, and f-butyl groups substituted for methyl at the 2- and 
4-positions and with phenyl, chlorine, and bromine substituted for hydrogen at the 
3-position of the parent 2,4-pentanedionates. Electronic polarization changes caused 
by substitution were found not to depend on the nature of the central metal. In 
contrast, the corresponding changes in atomic polarization (the difference be- 
tween the total and electronic polarization) upon substitution were found to 
depend on the metal. Substitution of phenyl for hydrogen at the 3-position pro- 
duces a smaller increment of electronic polarization than does such substitution at 
the 2,4-positions. The opposite effect was found for the atomic polarizations upon 
such substitution. Electronic polarization increments upon substitution of chlorine 
and bromine for hydrogen at the 3-position indicate that these halogens in the 
complexes have the character of being bonded to an aromatic ring, especially, so 
for bromine. 



Chemistry 179 

The Isolation of a New Denaturant-stable Protease from a Commercial Protease 
Preparation, Pronase. Wanda J. Wills and Eric R. Johnson, Department of 
Chemistry, Ball State University, Muncie, Indiana 47306. Pronase, a commer- 
cially available mixture of proteolytic enzymes, has been shown to contain two 
protease components that are stable and active in 6.0 M guanidinium chloride (W. 
M. Awad, Jr., et al. (1972) Proc. Nat. Acad. Sci. (USA) 69 2561-2565). During the 
purification of these two stable proteases from Pronase, a third component was 
isolated that survived incubation with the other two stable proteases for one 
week in 6.0 M guanidinium chloride. This newly isolated protease was found to 
possess esterase activity againt N-a-acetyl-L-tyrosine ethyl ester both in the 
absence and presence of 6.0 M guanidinium chloride, suggesting that this protease 
is also stable and active in the presence of denaturant. This new stable protease 
demonstrated chromatographic behavior on carboxymethylcellulose which was 
significantly different from that exhibited by previously isolated stable proteases. 
Polyacrylamide gel electrophoresis at pH 4.3 of the newly isolated stable protease 
resulted in a single band of substantially different electrophoretic mobility than 
the other stable Pronase components. 



Interactions of Various Homopolypeptides 
with Human Platelet-Rich Plasma Suspensions 

Barth H. Ragatz 

Fort Wayne Center for Medical Education, 

Indiana University School of Medicine, 

Fort Wayne, Indiana 46805 

Introduction 

A number of reports indicate that homogeneous polymers of L-lysine of 
various molecular weights will induce aggregation in stirred suspensions of human 
platelet-rich plasma (PRP) (2, 3, 4, 6, 7, 12). This aggregation phenomenon is concen- 
tration dependent (4, 7), molecular weight (size) independent in the range 
2,800-194,000 daltons (6, 12), positive charge dependent (5, 7), and is blocked by cer- 
tain naturally occurring polymers (heparin, chondroitin sulfate) with net negative 
charge (2, 7). There are conflicting reports as to whether the aggregation is calcium 
dependent (3, 4), plasma cofactor dependent (4), and whether it can be blocked by 
inhibitors of ADP induced aggregation or inhibitors of the platelet release reaction 
(2,3). 

The goals of this project are to expand the understanding of the polymer- 
platelet interaction at these several levels: (a) What are the interactions of other 
polymers (cationic, anionic) with PRP? (b) Are there smaller molecules (similar in 
monomer conformation) which can competitively saturate polymer receptor sites? 
(c) Are there cooperative interactions between polymer receptors and other recep- 
tors on the platelet surface? 

Materials and Methods 

Freshly collected whole blood from normal medication-free human subjects 
are rapidly mixed with one-tenth volume of 3.2% sodium citrate. Platelet-rich 
plasma (PRP) was prepared by differential centrifugation and the platelet count was 
adjusted to approximately 400,000mm 3 by dilution with 0.155 M NaCl. All 
homopolypeptides were obtained from either Sigma Chemical Company (St. Louis, 
Missouri) or from Miles Laboratories (Elkhart, Indiana) and were diluted in 0.155 M 
NaCl and adjusted to pH 7.0. Polymers utilized include Miles low molecular weight 
(MW = 14,000) poly-L-lysine (LPL); Sigma high molecular weight (MW = 165,000) 
poly-L-lysine (HPL); Sigma poly-L-arginine (MW = 14,000) (PA); Sigma poly-L- 
histidine (MW = 15,000) (PH); Sigma poly-L-glutamic acid (MW = 38,000) (PG); and 
Sigma poly-L-tyrosine (MW = 78,000) (PT). Platelet aggregation was studied by the 
turbidimetric method of Born (1) in a Payton dual channel aggregometer. Samples 
of PRP (0.5 ml) were placed in cuvettes within the aggregometer and were 
stimulated after 30 seconds by addition of 0.05 ml polypeptide solutions. In other 
experiments, 0.5 ml PRP was incubated initially with 0.05 ml of Sigma L-lysine or 
Sigma L-arginine, and challenged with an appropriate polymer (0.05 ml) 2-5 
minutes later. Finally, some PRP samples (0.5 ml) were incubated with LPL (0.05 
ml) initially and stimulated by Sigma adenosine - 5' - diphosphate (ADP) additions 
(0.05 ml) from 30 seconds to 6 minutes later. 

Results 

A typical example of platelet aggregation induced by a poly-L-lysine prepara- 
tion is shown in Figure 1. The extent of aggregation is seen to be polymer concen- 

180 



Chemistry 181 



A. HPL (O.Olmg/ml) 

B. HPL (O.lmg/ml) 

C. HPL (l.Omg/ml) 



z 
o 

to 
to 

to 
Z 
< 

at 



O 




MINUTES 

FIGURE 1: Effect of additions of various concentrations of high molecular weight 
poly-L-lysine to stirred human platelet-rich plasma. 

tration dependent over a 100-fold concentration range. Similar concentration 
dependent results were obtained also for LPL over a 10-fold concentration range 
(0.1 mg/ml-1.0 mg/ml). Both experiments are in agreement with published data for 
poly-L-lysine preparations obtained in other labs (4, 7). 

Under similar conditions, the strongly cationic polypeptide, PA, induced a 
vigorous aggregation, while the weakly cationic polypeptide, PH, was without ef- 
fect on PRP for incubation periods up to 7 minutes. The strongly anionic polypep- 
tide, PG, and the weakly anionic polypeptide, PT, were both devoid of inducing ag- 
gregation or producing shape changes in platelets during incubation periods of 5-7 
minutes 

Figure 2 shows that incubation of PRP with L-arginine causes no measurable 
turbidimetric change. Furthermore, if PA is added after incubating PRP with 
L-arginine for 3 minutes, the PRP sample is almost completely aggregated without 
any delay period. Analogous results have been obtained using PRP plus L-lysine, 
and PRP plus L-lysine plus HPL combinations. 

At this point in time, data have been obtained which indicate that LPL and 
ADP interact together in an additive manner in bringing about platelet aggrega- 
tion. Figure 3 shows that PRP incubated with a suboptimal concentration of LPL 
can be stimulated to aggregate additionally if ADP is added 30 seconds to 6 
minutes later. 



182 



Indiana Academy of Science 



A. PA (l.Omg/ml) 

B. L-Arginine(1.0mg/ml)+PA(1.0mg/ml) 3' Later 



o 




2 4 6 

MINUTES 

Figure 2: Examination of L-arginine as a potential competitive inhibitor ofpoly-L- 
arginine induced aggregation of human platelet-rich plasma. 



A. LPL (O.lmg/mll+ADPflxlO^M) 30" Later 

B. LPL (aimg/ml)+ADP(lxlO- 4 M) 2 Later 

C. LPL (0.1mg/ml)+ADP(lxlO- 4 M) 6' Later 




MINUTES 
Figure 3: Effect of time dependent additions of adenosine- 5 '-diphosphate to stirred 
human platelet-rich plasma suspensions pre treated with low molecular weight poly- 
L-lysine. 



Chemistry 183 

Discussion 

Present results for poly-L-lysine polymers are in agreement with those of 
other investigators; namely, that polymers in the molecular weight range 14,000 
- 165,000 are effective in inducing a dose dependent aggregation of PRR HPL seemed 
to be more effective at lower concentrations than LPL in producing aggregation. 
Literature reports suggest that poly-L-lysine samples as small as molecular 
weight 2,500 - 4,000 are able to stimulate platelet aggregation (4, 6, 7, 12). A low 
molecular weight poly-L-arginine, PA (MW = 14,000) is able to stimulate aggrega- 
tion, but a poly-L-histidine sample, PH, of similar molecular size (MW = 15,000) is 
devoid of effect. The anionic polymers, PG and PT, are also both without effect on 
PRP. These results obtained for five kinds of polymers suggest that the proag- 
gregatory effect is limited to poly cations only; hence, it is probably caused by 
electrostatic interaction between polymer and platelet surface, rather than by ac- 
tion at a receptor. That PA is stimulatory and PH is not suggest that charge den- 
sity and steric factors have some importance in determining the platelet-polymer 
interaction and that monomer structure is not a specific determinant. 

The importance of significant positive charge density on poly-L-lysine 
samples is supported also by the observation that succinylation of the terminal ep- 
silon amino groups of such polymers abolishes proaggregatory activity (7). Similar 
abolition of activity occurs after D-galactose is covalently coupled with poly-L- 
lysine epsilon amino groups (5). 

That the interaction between platelets and certain polycations is principally 
electrostatic is also supported by the following literature reports: (a) Natural 
polyanions (heparin, chondroitin sulfates) form complexes with polylysine and 
block the polymer aggregating effect (2, 7) and (b) Even D- or D, L- polylysine 
samples induce aggregation suggesting that polymer conformation is not an im- 
portant determinant of functionality (4). 

Present results also indicate that large concentrations of either L-lysine or 
L-arginine are without direct effect on platelets, and these amino acids are unable 
to occupy any discrete cationic binding sites on platelet membrane. Thus, the 
amino acids are unable to abolish the effect of either poly-L-lysine or poly-L- 
arginine. It would not be expected that the initial monomer concentrations added 
would be significantly lowered by metabolic events taking place in the platelet 
suspensions within the incubation periods used during these experiments. Jenkins 
et. al previously reported that L-lysine was without direct effect on PRP and had 
no influence on poly-L-lysine induced aggregation (2). Another literature report, 
however, related to the effect with added L-arginine stated that large concentra- 
tions of this amino acid inhibited collagen cross linking and multimer formation, 
rendering the fibrous protein inactive for aggregation of platelets (11). The lack of 
binding or interaction of basic amino acids with platelet membrane was also ex- 
tended by Rennert et. al. who reported that L-arginine and the urea cycle in- 
termediate, L-ornithine, were both without effect on platelet aggregation induced 
by ADP, epinephrine, or ristocetin (9). 

At present, our experiments have shown a cooperative interaction between 
LPL and ADP in induction of platelet aggregation. Massini et. al also observed 
that ADP added to PRP 30 seconds after addition of poly-L-lysine (MW = 50,000) 
enhanced aggregation (4). These effects may be easier to observe with smaller 
polymers as used in the present study or in the Massini et. al report since large 
molecular weight polymers may obstruct access of ADP or other inducers to 
specific platelet receptor sites. As seen in Figure 3 from the similar slopes and max- 



184 Indiana Academy of Science 

ima of the various curves, the interaction of ADP and LPL with platelets appears 
to be additive. These results are compatible with the hypothesis that cationic 
polymers attract platelets into close proximity by electrostatic interactions and 
that ADP acting at a specific unmasked receptor stimulates platelet aggregation 
and release of stored aggregating substances. 

Polycations have been shown to have other biological effects and additional in- 
teractions with plasma proteins which may be of importance to future studies with 
PRP. Vogel et. aL have used poly-L-lysines (MW range 6,900-72,000) to accelerate 
conversion of prothrombin to active thrombin by activated clotting factor X (13). 
Nossel et. aL reported that poly-L-lysine samples (MW range 4,000 - 100,000) inhibit 
the surface activation of Hageman factor (factor XII) (8). Secondary interactions of 
platelets with proteins of the blood coagulation or fibrinolysis systems can be 
eliminated by conducting experiments with platelets suspensions washed free of 
plasma proteins. Finally, Ryser, reported that poly-L-ornithine samples (MW range 
45,000 - 200,000) or poly-L-lysine samples (MW range 3,000 - 225,000) increase cell 
permeability (measured by albumin uptake) in Sarcoma S-180 cell cultures (10). It is 
presently unknown if platelet permeability is modified by polycations. 

Future experiments in this lab will be directed at investigation of potential in- 
teractions of small cationic polymers with other platelet aggregation inducer recep- 
tor sites, e.g., epinephrine receptors. Small cationic agents which can serve as an- 
tagonists that will bind to platelet surface without producing aggregation and will 
block aggregatory effects of larger polymers will be sought. The potential clinical 
relevance of these experiments is obvious in view of the undesirable interactions 
between platelets and biological polymers of damaged arterial wall or synthetic 
polymers found in certain prosthetic heart valves. 

Acknowledgments 

The author wishes to acknowledge the encouragement and financial support 
of Dr. P. G. Iatridis for the project while the author was a faculty member of the 
Northwest Center for Medical Education, Gary, Indiana. 

Literature Cited 

1. Born, G. V. R. 1962. Aggregation of Blood Platelets by Adenosine 
Diphosphate and Its Reversal. Nature. 194:927. 

2. Jenkins, C. S. P., M. A. Packham, R. L. Kinlough-Rathbone and J. F. 
Mustard. 1971. Interactions of Polylysine with Platelets. Blood. 37:395-412. 

3. Massini, P. and E. F. Luscher. 1972. On the Mechanism by which Cell Contact 
Induces the Release Reaction of Blood Platelets; the Effect of Cationic 
Polymers. Thrombosis. 27:121-133. 

4. Massini, P., L. C. Metcalf, U. Naf and E. F. Luscher. 1974. Induction of 
Aggregation and of the Release Reaction in Human Platelets by Polylysine. 
Haemostasis. 3:8-19. 

5. Mester, M., B. Kraska, J. Crisba and L. Mester. 1976. Effect of Amadori- 
Type Sugar Derivatives of Poly-L-lysine on Platelet Aggregation. Haemost- 
asis. 5:115-119. 

6. Metcalf, L. C. and D. J. Lyman. 1974. The effect of Conformational Changes 
on the Blood Platelet Reactivity of Polylysine. Thrombosis Research. 
5:709-717. 

7. Mohammad, S. F., H. Y. K. Chuang and R. G. Mason. 1977. Roles of Polymer 
Size and 6-Amino Groups in Polylysine-Platelet Interaction. Thrombosis 
Research. 1:193-202. 

8. Nossel, H. L., H. Rubin, M. Drillings and R. Hsieh. 1968. Inhibition of Hage- 
man Factor Activation. J. Clin. Invest. 47:1172-1180. 



Chemistry 185 

9. Rennert, 0., B. Buehler, T. Miale and D. Lawson. 1976. Polyamines and 
Platelet Aggregation. Life Sciences. 19:257-264. 

10. Ryser, H. J. P. 1967. A Membrane Effect of Basic Polymers dependent on 
Molecular Size. Nature. 215:934-936. 

11. Simons, E. R. , C. M. Chesney, R. W. Colman, E. Harper and E. Samberg. 
1975. The Effect of the Conformation of Collagen on its Ability to Aggregate 
Platelets. Thrombosis Research. 7:123-139. 

12. Tiffany, M. L. and J. A. Penner. 1976. Polylysine Aggregation of Human 
Blood Platelets. Thrombosis Research. 8:529-530. 

13. Vogel, C. N., R. J. Butkowski, K. G. Mann and R. L. Lundblad. 1976. Effect 
of Polylysine on the Activation of Prothrombin. Polylysine Substitutes for 
Calcium Ions and Factor V in the Factor Xa Catalyzed Activation of Prothrom- 
bin. Biochem. 15:3265-3269. 



Fluoride Content of Common Foods 

James T. Streator, Pam Bertram, and Caryl L. Coy 
Manchester College, North Manchester, Indiana 46962 

Introduction 

Before many of the controversies surrounding fluoridation' 8,9 ' of drinking 
water can be discussed intelligently, recent data regarding fluoride levels in com- 
monly eaten foods needs to be available. The purpose of this study was to deter- 
mine fluoride levels in foods found in the local community. This allows reasonable 
estimates to be made of total daily fluoride intake. 

Determinations of fluoride content were made on over 180 different foods 
prepared in either the Manchester College cafeteria, local homes, local restaurants, 
or purchased from local stores. Also, the fluoride level in the local water supply 
was monitored daily for three weeks. 

Experimental 

A 5 to 25 gram sample of food was collected and sealed into a plastic bag and 
refrigerated for analysis as soon thereafter as possible. The solid or semi-solid 
samples were mixed in a blender to produce homogeneous samples. A portion was 
weighed to the nearest 0.01 gram and set to dry at 100°C for 24 hours and then 
reweighed for calculation of percentage water content. A 1.00 gram sample was 
weighed into a platinum crucible 1 two milliliters of 20 M NaOH added, the contents 
mixed and the sample left to dry for 2 hours at 150°C. At the end of this drying 
time, the sample was transferred to a cold muffle furnace; controls were set for the 
temperature to hold at 560 °C, and the sample fused for 3 hours. After fusion and 
cooling, 5 ml of H 2 was added to dissolve the melt. It was sometimes necessary to 
break up the melt with a stainless steel spatula and use boiling water for dissolu- 
tion. The solution was transferred to a styrofoam cup and the pH adjusted to ap- 
proximately 6, using 8 M acetic acid. This was done dropwise to slow the evolution 
of C0 2 from the carbonates that had formed. The solution was filtered through 
medium porosity filter paper into a 25 ml volumetric flask containing 1.5 grams of 
TISAB 2 powder. After dilution, this solution was put into a plastic container for 
storage. Liquids such as coffee or wine were treated as received with TISAB and 
stored in plastic containers. Fluoride levels were determined using an Orion 
Specific Ion Fluoride Electrode 3 . 

Since no standard foods with known fluoride levels were available, multiple 
samples were run on a number of foods. In all cases, duplicates agreed within ± 
5% at the .5 ppm 4 level, and a recovery of 90% of fluoride added to vegetable 
samples was found. Considering that some samples of the same food taken from 
different containers varied by a factor of two, the observed precision was quite 
acceptable. 

Discussion and Results 

The recent USDA listing (7) of recommended daily allowances (RDA) for 
fluoride indicates that from 1.5 to 4.0 mg of fluoride is the RDA. Previous studies 
(3,4,5) have found fluoride ingestion ranging from .18 to 5.4 mg. Questions have been 
raised concerning possible increases in fluoride levels in foods due to processing 
foods using fluoridated water or simply due to increased fluoride release from 
phosphate fertilizers(6). One previous study concluded saying ". . . the fluoride 

186 



Chemistry 187 

content of the different food items which constitute the diets for infants from 
birth to 6 months is sufficient and that additional fluoride supplements would not 
be required(lO)." 

In this study, we calculated the daily intake of fluoride in North Manchester 
would typically be in the range of 1.5 mg depending upon the foods eaten (Table 
1). The local water contains about .55 ppm of fluoride and a proposed addition of 
0.45ppm fluoride would add about .5 mg to the daily intake of many local people. 

Table 1. Six representative meals and their fluoride content 
Amounts of foods correspond to listings in standard nutritional tables (2) 

Meal Contents Total mg fluoride 

1 Orange juice, wheaties with whole milk, 2 slices white bread, 

2 cups coffee .44 

2 2 eggs, 2 slices whole wheat bread, grape jelly, grape 

fruit juice, 1 glass 2% milk .13 

3 Swiss cheese sandwich, vegetable soup, 2% milk, water .28 

4 Hamburger with tomatoes and a bun, pepsi, french fries, water .35 

5 Fruit cocktail, beef barbecue, green beans, white bread, wine- 

catawba red, coffee, water .41 

6 Cottage cheese, apple sauce, pork chop, lima beans, white 

bread, mashed potatoes, tea-constant comment, water .74 



Floride levels in meat baby foods were high and a single jar of Gerbers' 
chicken baby food 5 would supply about .34 mg of fluoride. It is quite possible that 
the concern of Wiatrowski et al. as indicated above might be justified. However, 
levels in baby orange and apple juices were found to be below our detection limits 
of .01 ppm. 

Only two of the wines tested, Lancers and Almaden Chenin Blanc, showed 
levels below .01 ppm. As indicated in table 2, the average fluoride content was 
moderately high. Two major wine producers were contacted and both 6 claim no ad- 
dition of fluoride to their wines. The addition of sodium fluoride to retard 
enzymatic action was practiced in the past (9) but is not currently done. Of interest 
is that both grape juice and grape jelly showed fluoride levels of .6 and .21 ppm, 



Table 2. Food Categories and Fluoride Levels - Units are mg of Fluoride 
per 1000 grams of food as eaten (ppm) 7 

CATEGORY NUMBER OF SAMPLES 

Vegetables 22 

Soups 17 

Meats 13 

Baby Foods 15 

Wine 15 

Yogurt 7 

Cereals (toasted) 5 

• nil means below the detection limits of 0.01 ppm. 



RANGE 


AVERAGE ppm F 


nil*-1.96 


.25 


nil 3.01 


.26 


nil-1.68 


.40 


nil 2.73 


.89 


nil 2.80 


.94 


nil .06 


.01 


.341.25 


.84 



188 Indiana Academy of Science 

respectively. Perhaps there is a significant concentration of fluoride in grapes and 
this shows up in the wine. This certainly would be an interesting area to research 
further particularly since sampling is such a delightful task. 

Vegetables showed relatively low fluoride levels with only brocolli showing 
significant levels (1.96 and .5 ppm). The canning liquid from two samples each of 
canned corn and green beans showed fluoride levels similar to those found in the 
local water supply although the corn and the beans in these cans had levels 5-10 
times lower than the liquids. It is quite probable that the canning liquid reflected 
the fluoride levels in the water used in the canning process. 

While coffee was found to provide no fluoride in addition to that in the water 
used for brewing, the same can not be said for tea. All solid tea samples showed 
high levels of fluoride. These would provide a range of fluoride levels in the tea as 
drunk since varying amounts of instant tea or variable brewing times can be used. 
But generally, tea was high in fluoride. One glass of instant tea could provide as 
much fluoride as one liter of water containing one ppm fluoride. 

Summary 

Significant amounts of fluoride can be obtained from foods eaten locally. 
However, no evidence was found that current fluoride levels are significantly dif- 
ferent from those found as far back as 1966(6). Foods eaten in other communities 
within this region of the country would probably show fluoride levels similar to 
those in North Manchester. However, communities with fluoridated water might 
add up to a milligram of fluoride to the total daily intake making this intake around 
2.5 mg of fluoride. 

Notes 

1. The procedure was a modification of that found in (1). 

2. Total Ionic Strength Adjustment Buffer purchased from Fisher Scientific Co. 

3. The electrode output was read on a Sargent Welch model LSX expanded scale 
pH meter. 

4. Levels of fluoride reported in this paper will all be ppm (mg/1) of fluoride ion 
or total milligrams of fluoride ion. The fluoride levels of all foods will be 
reported as ppm of fluoride ion in the food as eaten or total mg of fluoride in 
the sample of the food. 

5. A level of 2.73 ppm was found. For the 128 g contents of the jar, this would 
provide 0.34 mg of fluoride. 

6. Great Western and Taylor, both in Hammondsport, New York, were con- 
tacted. Both of these companies produce wine in New York and California. 

7. A complete listing of all foods and their fluoride content is available as a com- 
puter output from the first author. Data also may be obtained on a disk com- 
patible with an Apple II computer. 

Literature Cited 

1. Baker, R. L. 1972. Determination of Fluoride in Vegetation using the Specific 
Ion Electrode. Anal. Chem. 44:1326-1327. 

2. Church, C. F. and H. N. Church. 1975. Food Values of Portions Commonly 
Used. J.B. Lippincott, Phil. 12th Edition. 

3. Farkas, C. S. 1975. Total Fluoride Intake and Fluoride Content of Common 
Foods: A review. Fluoride. 8:98-105. 

4. Jerard, E. and P. Patrick. 1973. Summing of Fluoride Exposures. Inter. J. En- 
vir. Studies. 4:141-155. 



Chemistry 189 

5. Kramer, L. et al. 1974. Dietary Fluoride in Different Areas in the U.S. Am. J. of 
Clin. Nutr. 27:590-594. 

6. Marier, J. R. and D. Rose. 1966. The Fluoride Content of Some Foods and 
Beverages. J. of Food Science. 31:941-946. 

7. National Academy of Science. 1980. Recommended Daily Allowences. U.S. 
Govt. Printing Office, Wash. D.C. Table II. 

8. Rogot, E. et. al. 1978. Trends in Urban Mortality in Relation to Fluoridation 
Status. Am.J. of Epidemiology. 107:104-112. 

9. Waldbott, G. L. 1978. Fluoridation: the Great Dilemma. Coronado Press, 
Lawrence KS. 

10. Wiatrowski, E. et al. 1975. Dietary Fluoride Intake of Infants. Pediatrics. 
55:517-522. 



ECOLOGY 



Chairman: W. Herbert Senft 
Department of Biology, Ball State University, Muncie, Indiana 47306 

Chairman-Elect: Richard W. Greene 
Department of Biology, University of Notre Dame, Notre Dame, Indiana 46556 



ABSTRACTS 

Changes in Yellow Bass Growth Rates and Density During the First Ten Years of 
Its Establishment in Monroe Reservoir. Robert L. Ball, Indiana Department of 

Natural Resources, Avoca, Indiana 47420. Growth rates of yellow bass, 

Morone mississippiensis, in Monroe Reservoir have declined rapidly since the first 
native year class developed in 1969. At the third annulus, back-calculated growth of 
the 1969 year class averaged 26.2 cm, compared to 19.6 cm for the 1976 year class. 
Most of the loss of growth has occurred by the end of the second year of life. 
Growth during the first year of life prior to 1974 averaged 12.0 cm, while it dropped 
to an average of 9.7 cm in the following five years. Average growth increments dur- 
ing the second year of life decreased 2.1 cm during the same period. 

Growth in terms of annual length increments indicates that growth has not 
decreased steadily and is not inversely related to density of yellow bass, although 
density in terms of kg/ha. of yellow bass has increased over the 10 year study 
period. 

Although the size of yellow bass in the Monroe sportfishing catch was good 
initially (fish averaged 0.13 kg each in 1973 and 1974), the mean weight dropped to 
0.10 kg in 1979. Reduced weight and presence of a bacterial infection in the adult 
population of this species was followed by a reduction in sportfishing harvest of 
yellow bass in 1979. Limited food supply seems to be the immediate cause of poor 
growth. Reduction of growth of larger bluegill, and both reduced growth and reduced 
abundance of yellow perch in Monroe, indicate a dearth of small crustaceans and 
larval insects for which these three species appear to be competing in the open 
areas of the lake. Also, a lack of open water predators, such as white bass and 
walleye, may have contributed to the decline in yellow bass growth rates. 

Habitat, Food, and Life History Overlap in Temporary Pond Snails: Evidence for 
Competition. Kenneth M. Brown, Department of Biological Sciences, Indiana 

University-Purdue University at Fort Wayne, Fort Wayne, Indiana 46805. 

Ponds ranging from heavily wooded and temporary to open and permanent in 
nature were quantitatively sampled for a period of three years to study niche 
relationships among the guild of pulmonate snails inhabiting them. Aplexa hyp- 
norum and Helisoma trivolvis were specialists in, respectively, wooded tem- 
porary ponds, and open permanent ponds. Lymnaea elodes and Physa gyrina 
were more common in partially wooded, temporary ponds. Food selection experi- 
ments indicate that A. hypnorum specializes on detritus, and L. elodes on 
periphyton, but that P. gyrina and H. trivolvis are generalists. Overlap in life 
history patterns indicates that congeners should rarely co-exist, and field data 
supports this hypothesis. Finally, field manipulation experiments indicate that 
faster growing species, that are food specialists, are better competitors. 

190 



Ecology 191 

Detrital Dynamics Regulate Mosquito Production in Treehole Ecosystems. 

Stephen R. Carpenter and Durland Fish, Department of Biology, University of 

Notre Dame, Notre Dame, Indiana 46556. Production of the treehole mosquito 

Aedes triseriatus is intimately linked to the rate of decay of leaf litter. Scanning 
electron micrographs reveal that mosquito larvae consume fungal hyphae and 
bacteria from surfaces of decaying leaves. Abundance of microbes is strikingly 
reduced by mosquito feeding, suggesting intense competition among larvae for 
food. Although mosquitoes reduce the abundance of the microflora, they increase 
its rate of respiration and thereby increase the rate of production of microbial 
biomass. 

A laboratory comparison of mosquito production at five larval densities (5, 
10, 20, 40, and 80 larvae) in microcosms composed of 300 ml of water and 1 g of 
either sugar maple, black oak, or beech leaf litter demonstrated that maple leaves 
are superior, and oak leaves are inferior, producers of mosquitoes. Maple leaves 
decompose faster than oak and beech leaves. Decay rates of detritus increase with 
increasing larval density, presumably because thinning of the microflora by mos- 
quito feeding enhances the metabolism of the detritus. However, this effect is con- 
founded by heavy mortality of larvae at high densities. Density dependent mor- 
tality was more severe on oak and beech than on maple. As larval density in- 
creases, elapsed time until pupation increases while substantial decreases in 
survivorship, pupal weights, and production occur. Our results suggest that 
strong density dependent regulation of larval mosquitoes occurs in nature, and 
that competition for food is the means of this regulation. 

Limiting Nutrients and Primary Productivity. William Y. B. Chang, Great Lakes 

Research Division, University of Michigan, Ann Arbor, Michigan 48109. 

Lake Monroe is the largest reservoir in Indiana with a daily mean productivity of 
200 mgC/m 2 /day in an observed range from 26 mgC/m 2 /day to 714 mgC/m 2 /day. It 
is a medium soft reservoir; the acid combining capacity varies from 0.28 - 0.71 
meq./l with a mean slightly above 0.5 meq./l. The results of diurnal changes in 
major nutrients, the C, N, and P ratios, and bioassay experiments indicated that 
phosphorus is the major limiting nutrient on algal photosynthesis in this lake. Sur- 
face photo-inhibition may be used to indicate the sufficiency of light for the 
species of algae in the water. The low productivity in December, January, 
February, and early March can be attributed to light limitation due to low water 
transparency. 

Attrition of White Ash in Red Pine Plantations in Eastern New York. William 
Bliss Crankshaw, Department of Biology, Ball State University, Muncie, Indiana 

47306. White ash (Fraxinus americana) has a high mortality in Red pine (Pinus 

resinosa) plantations in glaciated areas of eastern New York. Ash reproduction, 
from seed disseminated by wind, is one of the most common woody plants occurr- 
ing in the red pine plantations. A high mortality of this ash occurs at the pole 
stage (4-6 inch d.b.h.). A positive correlation was determined between the percent 
slope and ash mortality in plantations with equivalent spacing of pines. 

Success of Bald Cypress Seedlings in the Drawdown Zone at Salamonie Reservoir. 

William Bliss Crankshaw, Department of Biology, Ball State University, Muncie, 
Indiana 47306. In the spring of 1974 over 1000 two year old bald cypress seed- 
lings were planted in the drawdown zone at Salamonie Reservoir on the Upper 
Wabash watershed. Plantings were made on sites which varied in vegetation 
covers, percent slope, exposure, and soil texture. An evaluation of bald cypress 



192 Indiana Academy of Science 

establishment was made in 1980. Approximately one third of the planted seedlings 
survived. The highest establishment rate occurred behind groves of black willows 
which minimized damage from driftwood and ice floes at high pool level. 

Effect of Habitat Productivity, Permanence, and Predation on the Life History of 
A Temporary Pond Snail. Dennis Devries, Department of Biological Sciences, 
Indiana University-Purdue University at Fort Wayne, Fort Wayne, Indiana 46805. 
Two temporary ponds and one permanent pond in northeastern Indiana sup- 
port populations of the pulmonate snail Lymnaea elodes. Physico-chemical 
parameters vary among ponds with overall mean temperature and dissolved 
oxygen levels the highest in the permanent pond. Periphyton growth rates are also 
higher in the permanent pond. Field rearing experiments with predators excluded 
show that survival, growth, and reproductive rates are highest in the permanent 
pond as expected if the populations are food limited. However, populations reach 
the highest density in one of the temporary ponds where minnows are absent. 
Hence, vertebrate predators may control populations in more permanent ponds, 
and productivity may be important in vernal ponds. 

Periphyton Productivity of Three Sample Sites along Juday Creek, St. Joseph 
County, Indiana. John S. Fezy, and Richard W. Greene, Department of Biology, 
University of Notre Dame, Notre Dame, Indiana 46556. Periphyton producti- 
vity is being measured at three different types of stream settings along Juday 
Creek, a first order stream which empties into the St. Joseph River in north- 
western Indiana. The sample sites include areas where the stream passes through 
agricultural land, residential and natural habitats. Glass slides are being used as 
artificial substrates in both horizontal and vertical position to monitor periphyton 
production. It is measured at 3 week intervals and is based upon ash weight. Physi- 
cal stream parameters and water chemical analysis are being conducted to deter- 
mine whether there are nutrient fluctuations through the seasonal changes and 
what part, if any, they may play in productivity in this particular aquatic 
ecosystem. Periphyton production has been greatest in the agricultural setting, 
while the natural habitat has exhibited the least. Correlation between biomass 
fluctuation, discharge and water temperature has been observed. The amount of 
water flowing over the cells is an important aspect of the productivity within a 
stream ecosystem, continuously supplying and removing nutrients and various 
waste products from the zone of diffusion about each cell. The same general trends 
have been noted for both horizontal and vertical substrates, however, more 
definite relationships have been observed on vertical substrates than on horizontal 
ones. 

An Updated Evaluation of Sampling Efficiencies of Overstory Sampling Methods. 

Stephen W. Fletcher, Environmental Science and Engineering, Inc., Gainesville, 

Florida 32604. In 1958, Lindsey, Barton, and Miles published a comparison of 

sampling efficiencies of various forest sampling methods, tested in an Indiana 
forest. A recent assessment of methods in southern Louisiana allowed an oppor- 
tunity to use similar evaluation methods for comparing sampling methods along 
wetland-upland gradients and to comment upon the applicability of their conclu- 
sions for other regions. 

Study sites involved wetland, upland, and intermediate areas in cypress- 
tupelogum, southern mixed hardwood, and oak-pine communities of various ages. 
Each of two study sites were divided into the three zones, and a tally was made of 
all trees within 10m by 10m grid units. Six quadrat variations and two plotless 



Ecology 193 

methods for overstory density and basal area, as well as two cover estimation 
methods, were evaluated. This evaluation was based upon sampling efficiency and 
accuracy as compared to the relative importance values from the tally results. 

Using the concepts of %SEM and "constant adequacy level", it was deter- 
mined that circular quadrats, plotless methods, and cover estimates all were more 
efficient than square or rectangular quadrats. Plotless methods, although efficient, 
were not as accurate in depicting true species composition, as measured by Soren- 
son's similarity index comparisons to the tally results. 

It is concluded that for most quantitative sampling programs, range-finder- 
measured 100m 2 circular quadrats (in conjunction with the Bitterlich variable 
radius method for basal area) are best suited in this region as well as in Indiana 
communities. However, when there is less need for statistically defensible data, 
simple cover estimates can produce virtually identical results with greater effi- 
ciency. In certain conditions of low species diversity or sufficient stand size, the 
plotless methods may be the best methods. Such conditions must be properly 
validated on a site specific basis if dependable results are to be obtained with the 
plotless methods. 

Macrophyte Induced Fluctuations of Water Chemistry in an East-Central Indiana 
Borrow Pit Lake. Paul A. Glander and Thomas S. McComish, Department of 
Biology, Ball State University, Muncie, Indiana 47306. Selected water chemis- 
try parameters were measured in a 3.2 ha borrow pit lake from June to October in 
1978 after preliminary evaluation in 1977. Data were collected at a macrophyte sta- 
tion in shallow water and at a control station void of macrophytes in deep water. 

Dense beds of Potomogeton pectinatus L. and P. pusillus L. dominated at the 
macrophyte station. Dry weight macrophyte biomass, including associated Auf- 
wuchs, peaked in July but was gone by August. 

Photosynthesis at the macrophyte site accounted for elevated dissolved 
oxygen, and biomass was positively correlated with total dissolved oxygen. 
Respiration over night was positively correlated with biomass and accounted for 
lower morning dissolved oxygen. Photosynthesis increased pH and respiration 
lowered pH over night on most sample dates but the relationship was not signifi- 
cant. 

Macrophyte biomass had little influence on daily total alkalinity, however, 
there was a positive correlation with seasonal levels. Photosynthesis increased 
evening COJ alkalinity and decreased HC0 3 alkalinity. Morning levels of COJ 
alkalinity were lowered due to respiration while HC0 3 alkalinity increased. Macrop- 
hyte biomass was positively correlated with COg alkalinity and negatively cor- 
related with HCO3 alkalinity. 

The summer water chemistry dynamics of the lake were clearly regulated by lit- 
toral submerged aquatic macrophytes and associated Aufwuchs. These results 
demonstrate the impact of dense aquatic macrophytes on general water chemistry 
of shallow lentic systems in central Indiana. 

A Comparison of the Effect of Aluminum on a Single Species Algal Assay and 
Indigenous Community Algal Toxicity Bioassay. Joanne M. Payton and Richard 
W. Greene, Department of Biology, University of Notre Dame, Notre Dame, 

Indiana 46556. Since the enactment of the Toxic Substances Control Act 

(TSCA, PL 94-469) there has arisen a definite need for developing methods to 
evaluate the effects of toxic substances on aquatic organisms relative to the predic- 
tion of the effects on natural populations. This study simultaneously examines the 



194 Indiana Academy of Science 

effects of varying concentrations of aluminum at pH 5.0 on Scenedesmus bijgua, 
and on the indigenous phytoplankton community from Fishtrap Lake, LaPorte, 
Indiana. Experiments with algal assays have shown that there is a marked 
decrease in cell growth as aluminum concentrations increase from 500 ug/1 to 1000 
ug/1 Al. In the case of the indigenous phytoplankton community assay, plots of 
diversity versus time exposed to aluminum show an initially higher diversity index 
with a higher aluminum concentration. The plots also indicate a more dramatic 
decline in diversity as time progresses in the assays containing 1000 ug/1 Al. 

Growth and Phosphorus Uptake as a Function of Temperature in the Colonial 
Green Alga Volvox globator L. Ken Roberts and W. Herbert Senft II, Ball State 
University, Muncie, Indiana 47306. The growth of Volvox globator L. was mea- 
sured at five different temperatures and nine different phosphorus concentrations. 
Growth rate was hyperbolically related to phosphorus concentration for all 
temperatures using a Monod growth model. Optimal growth rates were dependent 
upon temperature and varied from 0.20 div/day (T = 10°C) to 1.17 div/day (T = 
20°C). No growth occurred at 30°C. Phosphorus uptake by V. globator was found 
also to be dependent upon external phosphorus concentrations and temperatures. 
Maximal uptake occurred at 30°C. The lack of growth of this algae at 30°C, there- 
fore, is not caused by physiological restrictions on phosphorus uptake. 

The Small Mammals of Spencer County, Indiana. Robert K. Rose, Old Dominion 

University, Norfolk, Virginia 23508. The small mammals of Spencer County, 

Indiana were examined during 4 studies conducted on a 1498 ha site near Rockport 
from 1973 through 1977, and briefly at nearby locations in late 1977. In all, 7 forest 
and 8 oldfield plots were studied on the Rockport site. Oldfields had both more 
species and more individuals. Prairie voles predominated in oldfields and white- 
footed mice in forests. Two concurrent snap- and pitfall-trapping studies revealed 
the presence of relatively large numbers of southeastern shrews, and in 1 study 
these shrews were the second most numerous species on the Rockport site. Pitfall 
traps yielded more species but fewer individuals than snap traps. The mammals 
collected in the county included 4 insectivores, 3 microtine rodents, 2 Old World 
murid rodents, white-footed mice, and meadow jumping mice. Analysis of barn owl 
pellets collected from a roost near the site provided an independent estimate of the 
composition of small mammal populations and also revealed the existence of the 
eastern cottontail and eastern woodrat. This study, which confirms the absence of 
the meadow vole in Spencer County, also provides new information about local 
abundance of southeastern shrews and southern bog lemmings and new distribu- 
tional records for meadow jumping mice and both masked and southeastern 
shrews. 

Trends in the Climatic Adaptations of Lichens. Robert Schwarzwalder, Jr., 

Department of Biology, Indiana University, Bloomington, Indiana 47405. Lichen 

ecology has traditionally shown a distinct bias toward temperate and boreal 
ecosystems. The dearth of research on subtropical and tropical ecosystems has 
resulted in unwarranted generalizations and an underestimation of the complexity 
of the lichen symbiosis. In an investigation of corticolous epiphytes in southern 
Louisiana I discovered variations in horizontal zonation patterns on isolated tree 
boles, microhabitat utilization and niche specialization which differed from pat- 
terns noted in temperate and boreal lichen studies. Personal observations on the 
microhabitat preferences of Candelaria concolor (Dicks.) Stein., a minutely foliose 
lichen common throughout the continental United States, and a French study of 
climatic influences on corticolous epiphyte distributions in Java indicate decreas- 



Ecology 195 

ing horizontal zonation and increasing microhabitat specialization with decreasing 
latitude. Latitudinal distribution also correlates with the growth form of lichens: 
fruticose forms being more abundant in cooler climates; crustose lichens, in 
warmer climates. Like other organisms, lichens display an increased diversity in 
form and niche specialization in tropical and subtropical climates. Variations in 
lichen growth appear to be due to differences in illumination, temperature, 
humidity and precipitation. Further study of these variations will expand and may 
reshape our understanding of the lichen symbiosis. 

Seasonal, Spatial and Developmental Variability of Benthic Macroinvertebrates 
in a Northern Indiana Stream. Bruce W. Schwenneker and Ronald A. Hellen- 
thal, Department of Biology, University of Notre Dame, Notre Dame, Indiana 
46556. Variability in the seasonal and spatial distribution of 29 benthic macro- 
invertebrate taxa from a riffle area of Juday Creek, a first order tributary of the 
St. Joseph River, was evaluated from November 1979 to May 1980. Five samples 
along each of three transects in an apparently uniform riffle area with an average 
velocity of 2.3 m/sec. were collected during November, January, April and May 
using a Hess Sampler. Organisms were separated from the substrate using sugar 
flotation, and were identified, sorted by instar, size or stage, and counted. 
Samples from each transect and sampling date were compared using analysis of 
variance, and components of variation attributable to location and date were 
estimated for each taxon. Season, stage, and location were found to affect both the 
densities of organisms and sample variability. Late instars of Chimarra obscura 
(Walker) (Trichoptera: Philopotamidae) had sample standard deviations which 
ranged from 6.8 to 0.53 on different dates. Sample variances for this species also 
varied along transects where samples taken near the center of the riffle had 
standard deviations as much as 4 to 8 times greater than those collected closer to 
the riffle margins. Sample variability also differed among life cycle stages and in- 
stars of the same species. Pupae of Symphitopsyche sparna (Ross) had a coeffi- 
cient of variation more than 3 times larger than last instar larvae. Since sample 
variability directly affects the assumptions and power of statistical tests, 
excessive and inconsistent sample variation complicates sampling strategies, 
experimental designs, and analyses. In studies of benthic communities employing 
secondary production or diversity measurements these complications may be pro- 
found. A preliminary sampling program can be used to identify and to estimate 
important components of sample variance. These estimates may be used in the 
development of efficient and effective sampling schemes. 

Spatial Patterning of a Volvox Globator L. Population in a Northern Minnesota 
Pond. W. Herbert Senft II and Arthur J. White, Department of Biology, Ball 
State University, Muncie, Indiana, 47306. The spatial distribution of the col- 
onial green alga Volvox globator L. is remarkably non-uniform in nature. Detailed 
sampling of a small, northern Minnesota pond in Lake Itasca State Park revealed a 
distinct clumping and aggregating pattern that is not related to wind mixing. The 
abundance of V. globator colonies in the water column Varied tremendously rang- 
ing from colonies/ml to a high of 17.5 colonies/ml. Dense aggregations of V. 
globator colonies were always associated with shallow water (<0.50 m) and areas 
of macrophyte abundance. In particular, the densest concentration of colonies was 
found in shallow water (<0.40 m) directly above a large mat of aquatic moss. It is 
not known whether macrophytes produce chemical changes in the water column 
necessary for Volvox growth or simply provide a structural retreat for the algal 
colonies. It is certain, however, that any quantitative samplings of Volvox popula- 



196 Indiana Academy of Science 

tions in small ponds must take into account the complex distributional patterns of 
these algae. 

Relationships of Algal Trophic State Indices in Indiana Lakes and Reservoirs. 

Patrick F. Sullivan and Stephen R. Carpenter, Department of Biology, Univer- 
sity of Notre Dame, Notre Dame, Indiana 46556. A comparison of 14 trophic 

state indices was based on 42 phytoplankton samples from 22 Indiana lakes and 
reservoirs. Trophic state indices included: Nygard's Myxophycean, Chlorophy- 
cean, Euglenophycean, diatom, and compound indices; Palmer's genus and species 
indices; average diversity (H'); number of taxa; maximum diversity; cell concentra- 
tion; evenness; mean number of taxa/ml; and concentration of the most abundant 
taxon. We used principal components analysis to determine the patterns of indices 
in sample space and samples in index space. Distortion due to curvature ("the 
horseshoe effect") was corrected by projecting points onto tangents of the arc of a 
quadratic fit to the ordination results, evidently a new method of linearizing curved 
ordination spaces. 

Average diversity explained about 80% of the total inter-lake variation. Peak 
average diversity occurred in mesotrophic samples, and average diversity 
decreased toward both oligotrophic and eutrophic ends of the gradient. Because 
average diversity varies non-monotonically with degree of eutrophication, com- 
parisons based on average diversity alone can yield misleading conclusions about 
relative trophic state. On the other hand, classical indices of trophic state 
(Nygard's, Palmer's, number of taxa, and cell concentration) appear to be 
monotonically related and yield more or less convergent results. We suggest that a 
species list (from which Nygard's and Palmer's indices are calculated) and a simple 
measurement of cell concentration provide a relatively reliable assessment of the 
trophic state of the phytoplankton. Little additional information about trophic 
state is obtained from the considerable labor involved in the species-specific cell 
counts needed to calculate average diversity. 



Fishes Reported from Waters 
of the Hoosier National Forest 

Forrest C. Brown and Thomas S. McComish 

Department of Biology, Ball State University 

Muncie, Indiana 47306 

Introduction 

McComish and Brown (4) reviewed all available literature prior to 1980 on 
scientific fish collections in waters of the Hoosier National Forest area. A com- 
prehensive species list was produced with appropriate life history notes and 
distribution maps for fishes. The review provides valuable insight to the species 
of fishes associated with waters of the Forest and will allow future comparative 
evaluation of the fish fauna with relative ease. 

Study Area 

The Hoosier National Forest is located in the Crawford and Norman uplands 
of nine southern Indiana counties. The southern forest area extends northward 
from the Ohio River through portions of Perry, Crawford, Orange, eastern Dubois, 
eastern Martin, and western Lawrence counties. A separate, smaller northern area 
includes northeastern Lawrence, northwestern Jackson, southwestern Brown, and 
southeastern Monroe counties. The northern area is bordered on the west by the 
Monroe Reservoir and on the north by Brown County State Park. 

Streams associated with the Forest are part of the Mississippi system, drain- 
ing into the Wabash River Basin in the northern and central areas and the Ohio 
River Basin in the southern area. Four major streams are included in the Wabash 
River Basin: Salt Creek, East Fork White River, Lost River, and Patoka River. The 
Ohio River drainage includes two major streams: Middle Fork Anderson River and 
Little Blue River. 

A wide variety of stream habitats in the Forest allows for the considerable 
diversity of fishes reported. This stream variety ranges from small, clear head- 
waters to deep, mature, and turbid rivers. Collections are also reported from semi- 
lentic stream impoundments ranging from small to large reservoirs. 

Collection Sites and Species at Sites 

A total of 87 collection sites were identified both within and nearby the 
exterior boundaries of the Forest (1), including: 27 from the East Fork of the White 
River, 11 from the Lost River (including Springs Valley Lake), 11 from the Patoka 
River system (prior to the impoundment of Patoka Reservoir), 15 from the Middle 
Fork of the Anderson River (including Indian Lake, Lake Celina, Tipsaw Lake, and 
Saddle Lake), 14 from the Little Blue River (including English Reservoir), and one 
each from Ohio River embayments at Deer Creek and Oil Creek. 

A total of 104 species were reported from these collection sites. The number of 
species from individual sites were as follows by system: 1 to 34 in Salt Creek 
(including Monroe Reservoir), 1 to 18 in the East Fork of the White River, 6 to 27 in 
the Lost River, 4 to 18 in the Patoka River, 8 to 21 in the Middle Fork Anderson 
River, 4 to 21 in the Little Blue River and 13 and 11 respectively, in Ohio River 
embayments at Deer Creek and Oil Creek. In terms of major stream systems. Salt 
Creek had the greatest number of species while the East Fork White collections 
yielded the fewest. 

197 



198 



Indiana Academy of Science 



Fish Taxa 

In addition to 104 species actually collected, 4 species not collected in stream 
surveys but known from waters of the Forest (clarified later) were included in the 
total list of 108 species (1). This total fish fauna list represents 48 genera in 21 
families of 13 orders (Table 1). The five largest families with numbers of species 
included: Cyprinidae (minnows) - 29 species, Percidae (perches) - 16 species, Cen- 
trarchidae (sunfishes) - 14 species, Catostomidae (suckers) - 13 species, and 
Ictaluridae (catfishes) - 10 species. A total of 82 species, 76 percent of all 108 
species, were represented in these five families. The remaining 26 species 
represented 16 families having 1 to 4 species. 

Occurrence by Species 

The fishes were grouped on the basis of frequency of occurrence at sites in the 



Table 1. Numbers of orders, families, genera, and species of fishes known from 
waters of the Hoosier National Forest area (k). 



Order/Family 



Genus Species 

(Genera) 



Petromyzontiformes 

Petromyzontidae (lampreys) 
Acipenseriformes 

Acipenseridae (sturgeons) 
Semionotiformes 

Lepisosteidae (gars) 
Amiiformes 

Amiidae (bowfins) 
Anguilliformes 

Anguillidae (eels) 
Clupeiformes 

Clupeidae (herrings) 
Osteoglossiformes 

Hiodontidae (mooneyes) 
Salmoniformes 

Esocidae (pikes) 
Cypriniformes 

Cyprinidae (minnows) 

Catostomidae (suckers) 
Siluriformes 

Ictaluridae (catfishes) 
Percopsiformes 

Amblyopsidae (cavefishes) 

Aphredoderidae (pirate perches) 

Percopsidae (trout-perches) 
Atheriniformes 

Cyprinodontidae (killifishes) 

Atherinidae (silversides) 
Perciformes 

Cottidae (sculpins) 

Percichthyidae (temperate basses) 

Centrarchidae (sunfishes) 

Percidae (perches) 

Sciaenidae (drums) 



Totals 



Ecology 



199 



Forest area (Table 2). Frequency categories included: very common (>40 sites), 
common (11-40 sites), uncommon (2-10 sites), and rare (1 site). 

Five species were very common and generally distributed among collection 
sites: Lepomis megalotis (longear sunfish) - 68 sites, Lepomis macrochirus (bluegill) 
- 53 sites, Micropterus salmoides (largemouth bass) - 50 sites, Pimephales notatus 
(bluntnose minnow) - 45 sites, and Lepomis cyanellus (green sunfish) - 44 sites. 

There were 38 common species. Major representatives (>20 sites) of this 
group included: Campostoma anomalum (stoneroller) - 34 sites, Notropis cornutus 
(common shiner) - 32 sites, Lepomis gulosus (warmouth) - 31 sites, Catostomus com- 



Table 2. Fishes reported (4) from Hoosier National Forest waters, number of 
collection sites, and frequency category (with sites in parentheses). 









Frequency Category/(No. Sites) 


Family/Scientific Name 


Common Name 


Total 
Sites 


Very 
Common Common Uncommon Rare 
(>40) (11-40) (2-10) (1) 


Petromyzontidae 








Ichthyomyzon bdellium 
Ichthyomyzon castaneus 
Ichthyomyzon unicuspis 
Lampetra aepyptera 


Ohio lamprey 
Chestnut lamprey 
Silver lamprey 
Least brook lamprey 


1 
3 
2 

1 


X 
X 
X 

X 


Acipenseridae 








Scaphirhynchus platorynchus 


Shovelnose sturgeon 


1 


X 


Lepisosteidae 








Lepisosteus osseus 
Lepisosteus platostomus 


Longnose gar 
Shortnose gar 


4 
2 


X 
X 


Amiidae 








Amia calva 


Bow fin 


3 


X 


Anguillidae 








Anguilla rostrata 


American eel 


1 


X 


Clupeidae 








Alosa chrysochloris 
Dorosoma cepedianum 


Skipjack herring 
Gizzard shad 


9 
24 


X 
X 


Dorosoma petenense 


Threadfin shad 


1 


X 


Hiodontidae 








Hiodon tergisus 


Mooneye 


1 


X 


Esocidae 








Esox americanus vermiculatus 


Grass pickerel 


27 


X 


Esox lucius 


Northern pike 


2 


X 


Cyprinidae 








Cyprinus carpio 
Phoxinus erythrogaster 
Hybopsis amblops 
Hybopsis storeriana 


Carp 

Southern redbelly dace 

Bigeye chub 

Silver chub 


24 
9 

1 
1 


X 

X 

X 
X 


Hybopsis xpunctata 
Nocomis biguttatus 
Semotilus atromaculatus 


Gravel chub 
Hornyhead chub 
Creek chub 


1 

2 

28 


X 
X 
X 


Ericymba bucatta 
Phenacobtus mirabilis 


Silverjaw minnow 
Suckermouth minnow 


12 
2 


X 

X 



200 


Indiana Academy of Science 






Table 2. Continued 














Freq 


uency Category/(No. Sites) 






Very 






Family/Scientific Name 


Common Name 


Total Common 


Common 


Uncommon Rare 






Sites (>40) 


(11-40) 


(2-10) (1) 


Pimephales notatus 


Bluntnose minnow 


45 X 






Pimepkales promelas 


Fathead minnow 


9 




X 


Pimephales vigilax 


Bullhead minnow 


9 




X 


Notemigonus crysoleucas 


Golden shiner 


16 


X 




Notropis amnis 


Pallid shiner 


1 




X 


Notropis ardens 


Rose fin shiner 


3 




X 


Notropis atherinoides 


Emerald shiner 


15 


X 




Notropis boops 


Bigeye shiner 


16 


X 




Notropis chalybaeus 


Ironcolor shiner 


1 




X 


Notropis chrysocephalus 


Striped shiner 


13 


X 




Notropis cornutus 


Common shiner 


32 


X 




Notropis emiliae 


Pugnose minnow 


4 




X 


Notropis photogenis 


Silver shiner 


3 




X 


Notropis rubellus 


Rosyface shiner 


2 




X 


Notropis shumardi 


Silverband shiner 


1 




X 


Notropis spilopterus 


Spotfin shiner 


9 




X 


Notropis umbratilis 


Redfin shiner 


23 


X 




Notropis volucellus 


Mimic shiner 


5 




X 


Notropis whipplei 


Steelcolor shiner 


15 


X 




Campostoma anomalum 


Stoneroller 


34 


X 




Catostomidae 










Ictiobus bubalus 


Smallmouth buffalo 


8 




X 


Ictiobus cyprinellus 


Bigmouth buffalo 


6 




X 


Carpiodes carpio 


River carpsucker 


8 




X 


Carpiodes cyprinus 


Quillback 


8 




X 


Carpiodes ve lifer 


Highfin carpsucker 


1 




X 


Erimyzon oblongus 


Creek chubsucker 


14 


X 




Moxostoma anisurum 


Silver redhorse 


1 




X 


Moxostoma duquesnei 


Black redhorse 


8 




X 


Moxostoma erythrurum 


Golden redhorse 


23 


X 




Moxostoma macro lepidotum 


Shorthead redhorse 


2 




X 


Catostomus commersoni 


White sucker 


30 


X 




Hypentelium nigricans 


Northern hog sucker 


15 


X 




Minytrema melanops 


Spotted sucker 


25 


X 




Ictaluridae 










Ictalurus catus 


White catfish 


1 




X 


Ictalurus me las 


Black bullhead 


10 




X 


Ictalurus natalis 


Yellow bullhead 


24 


X 




Ictalurus nebulosus 


Brown bullhead 


7 




X 


Ictalurus punctatus 


Channel catfish 


12 


X 




Pylodictis olivaris 


Flathead catfish 


6 




X 


Noturus eleutherus 


Mountain madtom 


1 




X 


No turns flavus 


Stonecat 


6 




X 


Noturus gyrinus 


Tadpole madtom 


8 




X 


Noturus miurus 


Brindled madtom 


4 




X 


9 
Amblyopsidae 










Aphredoderidae 










Aphredoderus sayanus 


Pirate perch 


20 


X 




Percopsidae 










Percopsis omiscomaycus 


Trout-perch 


1 




X 


Cyprinodontidae 










Fundulus notatus 


Blackstripe topminnow 


20 


X 





Ecology 



201 



Table 2. Continued 









Frequency CategoryANo. Sites) 








Very 






Family/Scientific Name 


Common Name 


Total 


Common 


Common 


Uncommon Rare 






Sites 


(>40) 


(11-40) 


(2-10) (1) 


Atherinidae 












Labidesthes sicculus 


Brook silverside 


14 




X 




Cottidae 












Cottus bairdi 


Mottled sculpin 


4 






X 


Cottus carolinae 


Banded sculpin 


8 






X 


a 

Percichthyidae' 












Morone mississippiensis 


Yellow bass 


4 






X 


Centrarchidae 












Micropterus dolomieui 


Smallmouth bass 


10 






X 


Micropterus punctulatus 


Spotted bass 


21 




X 




Micropterus salmoides 


Largemouth bass 


50 


X 






Ambloplites rupestris 


Rockbass 


15 




X 




Centrarchus macropterus 


Flier 


1 






X 


Lepomis cyanellus 


Green sunfish 


44 


X 






Lepomis gib bos us 


Pumpkinseed 


1 






X 


Lepomis gulosus 


Warmouth 


31 




X 




Lepomis humilis 


Orangespotted sunfish 


13 




X 




Lepomis macrochirus 


Bluegill 


53 


X 






Lepomis megalotis 


Longear sunfish 


68 


X 






Lepomis microlopkus 


Redear sunfish 


27 




X 




Pomoxis annularis 


White crappie 


14 




X 




Pomoxis nigromaculatus 


Black crappie 


10 






X 


4 
Percidae 












Perca flai'escens 


Yellow perch 


13 




X 




Stizostedion canadense 


Sauger 


1 






X 


Etheostoma blennioides 


Greenside darter 


11 




X 




Etheostoma caeruleum 


Rainbow darter 


14 




X 




Etheostome camurum 


Bluebreast darter 


1 






X 


Etheostoma flabellare 


Fantail darter 


25 




X 




Etheostoma nigrum 


Johnny darter 


29 




X 




Etheostoma spectabile 


Orangethroat darter 


12 




X 




Etheostoma tippecanoe 


Tippecanoe darter 


1 






X 


Percina caprodes 


Logperch 


16 




X 




Percina copelandi 


Channel darter 


2 






X 


Percina maculata 


Blackside darter 


11 




X 




Percina phoxocephala 


Slenderhead darter 


3 






X 


Percina sciera 


Dusky darter 


1 






X 


Percina shumardi 


River darter 


2 






X 


Sciaenidae 












Aplodinotus grunniens 


Freshwater drum 


18 




X 




TOTALS 






5 


38 


38 23 



1 Esox masquinongy ohioensis, Ohio muskellunge, reported in the Little Blue River 
system by sport fishermen through the 1960's. 

Amblyopsis spelaea, Northern cavefish, reported from cave explorations with 
records maintained by the U.S. Forest Service, Bedford, Indiana. 
3 Morone chrysops, white bass, stocked in Patoka Reservoir by the Indiana Depart- 
ment of Natural Resources. 

4 Stizostedion vitreum vitreum, walleye, stocked in Patoka Reservoir by the Indiana 
Department of Natural Resources. 



202 Indiana Academy of Science 

mersoni (white sucker) - 30 sites, Etheostoma nigrum (Johnny darter) - 29 sites, 
Semotilus atromaculatus (creek chub) - 28 sites, Esox americanus vermiculatus 
(grass pickerel) - 27 sites, Estheostoma flabellare (fantail darter) - 25 sites, 
Minytrema melanops (spotted sucker) - 25 sites, and Lepomis microlophus (redear 
sunfish) - 21 sites. 

Uncommon fishes included 38 species. Cyprinids (11 species), catostomids (6 
species), and ictalurids (6 species) were major uncommon fishes. 

Rare fishes of the Forest waters included 23 species. Fourteen of the rare 
species are particularly notable because they were reported only once over 25 
years ago and have not been reported since. Krumholz (3) reported in 1948 that he 
collected Dorosoma petenense (threadfin shad) and Hybosis storeriana (silver 
chub) in the Little Blue River near the Ohio River. Lampetra aepyptera (least 
brook lamprey) and Scaphirhynchus platorynchus (shovel nose sturgeon) were 
reported in 1955 by Gerking (2). Ten species were reported by Gerking in 1945 (1); 
Hidon tergisus (mooneye), Hybopsis amblops (bigeye chub), Hybopsis x-punctata 
(gravel chub), Notropis amnis (pallid shiner), Noturus cleutherus (mountain mad- 
tom), Percopsis omiscomaycus (trout-perch), Centrarchus macropterus (flier), 
Etheostoma camurum (bluebreast darter), Etheostoma tippecanoe (Tippecanoe 
darter), and Percina shumardi (river darter). 

A statement of clarification is necessary for the following four species not col- 
lected, but included in the total of 108 noted earlier. Morone chrysops (white bass) 
and Stizostedion vitreum vitreum (walleye) have been introduced in the Patoka 
Reservoir impoundment area. Subsequent investigations will establish their posi- 
tion and presence in the Forest waters fish fauna. Esox masquinongy ohioensis 
(Ohio muskellunge) was reported periodically in the Little Blue River system by 
sport fishermen, but it has never been collected in scientific surveys. Finally, the 
presence of Amblyopsis spelaea (northern cavefish) in the Forest waters has been 
established from cave studies. Cavefish records are maintained by the U.S. Forest 
Service, Bedford, Indiana. 

Summary 

The literature review revealed a total of 108 fish species reported from waters 
of the Hoosier National Forest in southern Indiana. Fishes were collected at 87 
defined sample sites with four species reported from other sources. The 108 species 
represented 21 families with 82 (76 percent) in only five families: Cyprinidae (29), 
Percidae (16), Centrarchidae (14), Catostomidae (13), and Ictaluridae (10). 

Five species were designated very common in distribution, occurring in at 
least half of the collection sites. Fishes occurring at 11 through 40 sites were con- 
sidered common and totaled 38 species. A total of 38 species were designated un- 
common in distribution and were sampled at 2 through 10 sites. Fish species con- 
sidered rare were collected at only one site. Of the 23 species designated as rare, 14 
species were collected at least 25 years ago making the current status of their 
distribution questionable. 

This manuscript is a brief synopsis of a report titled: Distribution and Life 
History Notes for Fishes of the Hoosier National Forest, Indiana by McComish 
and Brown in 1980 (4) as prepared for the U.S. Forest Service, Bedford, Indiana. 
This comprehensive report, available in Bracken Library, Ball State University, 
Muncie, Indiana, and the Indiana State Library, Indianapolis, Indiana, contains 
detailed and specific information on distribution and literature sources. 



Ecology 203 

This work provides basic information for recreation and research interests 
and a comparative baseline for future studies. Future sampling may reveal signifi- 
cant changes in fish fauna distribution due to introductions, past sampling deficien- 
cies, changes in habitat and water quality, and other factors. This comprehensive 
fish list provides a basis for decision-making in future management considerations 
and protection of the fishes and waters of the Hoosier National Forest, Indiana. 

Acknowledgment 

The authors wish to thank the U.S. Forest Service and Ball State University 
for funding in support of the project. Specific recognition is due H. E. McReynolds 
and K. Landes of the U.S. Forest Service, Bedford Indiana, for cooperation, sugges- 
tions, and assistance. 

Literature Cited 

1. Gerking, S. D. 1945. Distribution of the fishes of Indiana. Investigations of 
Indiana Lakes and Streams 3(1): 1-137. 

2. Gerking, S. D. 1955. Key to the fishes of Indiana. Investigations of Indiana 
Lakes Streams 4(2): 49-86. 

3. Krumholz, L. A. 1958. Second annual report of ORSANCO-Univ. Louisville 
Aquatic-Life Resources Project (mimeograph). 128 p. 

4. McComish, T. S. and F. C. Brown. 1980. Distribution and life history notes for 
fishes of the Hoosier National Forest, Indiana. Final Report 1978-1979 U.S. 
Forest Service Contract (mimeograph). 225 p. 



Fishes of Spicer Lake 

Clarence F. Dineen 
Department of Biology, Saint Mary's College, Notre Dame, Indiana 46556 

Introduction 

In a previous publication (2) on the open water plankton and benthos of Spicer 
Lake, Indiana; the acquisition, general description and the management of Spicer 
Lake Nature Preserve were discussed. Also, the significance of the floating mat 
area of the lake and the apparent small population of fishes were suggested as 
important aspects for future research. This study pertains to the fish population. 

Method 

The fishes were collected primarily by double-cone plastic minnow traps. Each 
trap was secured in position by a weight on the bottom and marked by a float. Some 
traps were lined with the fine mesh screen in order to catch the young of the year. 
Seines and dip nets operated from a boat were also used. Fishes were observed in the 
upper 0.5 m of water adjacent to the floating mat and in open areas of the mat. 
Breeding pairs of mudminnows and young were seen in depressions in the wooded 
floodplain in early spring. Repeated attempts with rod and tackle produced no 
fish. The open water of Spicer Lake was always quiet even on very windy days. 
However, no fish was ever seen "breaking water" while feeding on surface dwell- 
ing organisms. 

For quantitative data 8 to 12 double-cone traps were distributed equally 
around the lake adjacent to or in open areas of the inner edge of the floating mat. 
The Petersen estimate (Lincoln index) was used to determine the size of the popu- 
lation in 1980 (1). The distal, rounded edge of the anal fin was clipped for recap- 
ture identification. Some traps were baited with banana peel, peanut butter and 
commercial bait. 

Total length measurements were recorded in the field. Additional individuals 
were preserved and returned to the laboratory for measurements and stomach 
analyses to obtain food consumption data. 

Results and Discussions 

The Central mudminnow, Umbra limi (Kirtland) and the golden shiner, 
Notemigonus crysoleucas (Mitchell) inhabited Spicer Lake. The distribution of 
these species were limited to the floating mat area except in early spring when 
the mudminnow reproduced in microhabitats in the red maple floodplain which 
surrounds the lake. 

The mudminnow was the only species found in Spicer Lake in 1978 and 1979. 
Three winters, 1976-77, 1977-78 and 1978-79, with low mean temperatures coupled 
with record-breaking snow cover, 129.2, 172.0 and 101.3 total annual inches, 
respectively (9) perhaps resulted in a winter kill of other species. However, prior 
to this study the only records of fishes in Spicer Lake were the reports of 
fishermen and residents in the vicinity. They reported that several species of 
game fishes were taken from Spicer Lake in recent years. The mudminnow is 
tolerant to very low oxygen concentrations (3), consequently, a small population 
survived the previous severe winter conditions. Baiting the double-cone traps did 
not increase significantly the number of mudminnows caught. 

In the spring of 1980 the golden shiner was taken in the minnow traps and 

204 



Ecology 205 

observed in the floating mat area of the lake. The drainage of Spicer Lake is via 
the Dowling Creek outlet to the northeast which joins the Galien River in 
Michigan which then empties into the Saint Joseph River. The high water in the 
spring of 1980 in the Dowling Creek drainage permitted easy access for the golden 
shiner to Spicer Lake. The golden shiner inhabits vegetated waters of small 
streams and swamps, has wide ecological tolerance to many environmental fac- 
tors, and spawns by scattering adhesive eggs on vegetation (3, 8). Spicer Lake and 
the drainage system is a natural habitat for the golden shiner. 

The complex vegetative floating mat in Spicer Lake provided both protection 
from predators as well as an abundance of prey for the adult, carnivorous mud- 
minnows. The mudminnow predators were limited to some insect larvae, and as a 
minor food item in the diet, the green frog. Rana clamitans malanota. 

The breeding behavior of the mudminnow involves a shift of the population 
from stream or lake water to floodplains. The stimuli to spawn include the forma- 
tion of suitable microhabitats and a rise in the temperature of the water (6). Small 
depressions, microhabitats, in the floodplain around Spicer Lake supported large 
dipteran populations which are major food items for newly hatched mudminnows. 
In early spring of 1980 pairs of breeding mudminnows were common in the flood- 
plain on the east side of Spicer Lake. However, the extensive maple forested 
floodplain around the lake would support a very large population of mudminnows 
during the breeding season. 

Growth rings, annuli, are not formed in the scales of the mudminnow, thus age 
determinations are not feasible by a study of the scales. A study of otoliths is dif- 
ficult and the results are inconclusive (5). However, an analysis of the length fre- 
quency has been used by numerous researchers (4, 5, 6, 10). In this study total 
length measurements were used because the homoceral tail minimizes possible 
error and the dark pigmented caudal region inhibits standard length determina- 
tions. 

The mudminnows collected in Spicer Lake during this study ranged in total 
length from 12 to 140 mm. Smaller individuals were observed in the microhabitats 
in the floodplain during the breeding season. Young of the year travel from 
floodplain to the main body of water at total lengths ranging from 20 to 30 mm (5, 6, 

7). 

Sample collections of 200 individuals in late summer 1980 indicated 4 age 
groups when plotted in a histogram. The young of the year ranged in total length 
from 30 to 60 mm and the second year individuals from 60 to 80 mm. The numbers 
in each sample were represented about equally in these two age groups. A third 
group, three year, ranged from 80 to 95 mm. Less than 1.0% of the mudminnows 
collected obtained this age category. Those placed in the fourth year range, 110-140 
mm, were represented by less than 0.05 percent. 

The Petersen estimate (Lincoln index) with the margin of the anal fin clipped 
as a marker, resulted in populations estimates ranging from 8,000 to 12,000 mud- 
minnows in Spicer Lake in late summer of 1980. These are perhaps very conser- 
vative estimates. However, the small total number and the distribution of 
individuals in the four age groups indicate that the severe winter conditions, 
1976-79 perhaps reduced the total population of mudminnow in Spicer Lake. Future 
studies would be of interest to determine the possible growth of the mudminnow 
population. 

The food habits of the mudminnow in Spicer Lake, as indicated by stomach 



206 Indiana Academy of Science 

analyses were very similar to those studied in a small stream (6). The mudminnow 
was carnivorous, and size of food items was more important than the species of 
prey. Planktonic organisms, in particular cladocerans, were abundant in the 
stomachs of the young of the year mudminnows. Also, red mites and small annelids 
were common food items. Quantitative data on the size of the various invertebrate 
populations in the floating mat area were not established. However, the frequency 
of invertebrate taxa eaten by adult mudminnows was perhaps correlated directly 
to the abundance except for immature Odonata. Damselflies and dragonflies were 
extremely abundant in the floating mat area, however, few appeared in the 
stomach analyses. The most common prey were Diptera in the families Ten- 
dipedidae and Culicidae. Coleoptera, chiefly Dytiscidae, Hydrophilidae, Gyrinidae 
and Haliplidae were significant food items. Also, to a lesser extent, Hemiptera 
families were represented by Gerridae, Corixidae, Notonectidae and 
Belostomatidae. A few snails, leeches, mayflies and caddisflies were consumed by 
mudminnows. 

The golden shiners taken in Spicer Lake in 1980 consist of three distinct size 
groups within a size range, standard length, from 39 to 88 mm. Fifty-one percent of 
the 91 individuals collected ranged in size from 39 to 56 mm; 36 percent 60 to 66 mm 
and 13 percent 71 to 88 mm. The golden shiner inhabited the same area of the lake 
as the mudminnow. No golden shiners nor mudminnows were collected from the 
open water in the central portion of the lake. The intestinal tract of golden shiners 
contained fine detritus. 

Summary and Conclusions 

The Central mudminnow, Umbra limi, was the only species of fish collected 
from Spicer Lake in 1978-79. Three severe winters may have destroyed popula- 
tions of other species. Notemigonus crysoleucas entered the lake in 1980 via the 
Dowling Creek drainage system. The adults of both species of fishes inhabit the 
floating mat area and the mudminnow reproduced in the surrounding floodplain. 

Based on total length measurements the mudminnow population consisted of 
four age groups with a total range in size from 12 to 140 mm. The present popula- 
tion was judged to be in the range of 8,000 to 12,000 by using the Petersen mark- 
capture estimate. Food and shelter in the extensive floating mat, a lack of 
predators, as well as an abundance of microhabitats for reproduction indicated that 
the Spicer Lake ecosystem could support a much larger population of mud- 
minnows. The major food items of the young mudminnows were chiefly planktonic 
taxa; Cladocera was predominant. The adult consumed various invertebrate taxa 
in particular insects. 

The golden shiner population consisted of three distinct size groups within a 
total range in size, standard length, from 39 to 88 mm. The intestinal tract of the 
golden shiner contained only fine detritus. 

Literature Cited 

1. Caughley, G. 1977. Analysis of vertebrate populations. John Wiley & Sons. 
New York, New York 234 pp. 

2. Dineen, C. F. 1979. Plankton and benthos of Spicer Lake. Proc. In. Acad. Sci. 
50:173-179. 

3. Eddy, S. and J. C. Underhill. 1974. Northern fishes, with special reference to 
the upper Mississippi valley. Univ. of MN Press pp. 414. 

4. Jones, J. A. 1973. The ecology of the mudminnow, Umbra limi in Fish Lake 
(Anoka County, Minnesota). Ph.D. Thesis Iowa State University pp. 114. 



Ecology 207 

5. Peckham, R. S. 1955. Ecology and life history of the Central mudminnow, Um- 
bra limi (Kirtland). Ph.D. Thesis University of Notre Dame. pp. 71. 

6. Peckham, R. S. and C. F. Dineen. 1957. Ecology of the Central mudminnow 
Umbra limi (Kirtland). Proc. In. Acad. Sci. 55:223-231. 

7. Peckham, R. S. and C. F. Dineen. 1963. Development of the caudal fin in the 
Central mudminnow, Umbra limi (Kirtland). Copeia. 1963 IS): 586-588. 

8. Smith, P. W. 1979. The fishes of Illinois. University of IL Press, pp. 314. 

9. U. S. Department of Commerce. Local Climatological Data. South Bend, 
Indiana. 

10. Westermann, J. R. 1941. A consideration of population life-history studies in 
their relation to the problems of fish management research with special 
reference to the small-mouth bass, Micropterus dolomieu (Lacepede), the lake 
trout, Christivomer namaycush (Walbaum), and the mud minnow, Umbra limi 
(Kirtland). Ph.D. Thesis Cornell University, pp. 142. 

Acknowledgments 

A grant from Saint Mary's College supported this study of fishes. Also, I wish 
to thank the individuals who aided me with the field work: Mike Tanner, Richard 
Jensen, James Litton, David Sever, Joseph Bonadies and the Strickler boys, David 
and Bob. 



The Implications of Rotenone Eradication 

on the Fish Community of Eagle Creek in 

Central Indiana 

William L. Fisher 

Water Resources Laboratory, University of Louisville, 

Louisville, Kentucky 40292 

and 

James R. Gammon 

Department of Zoology, DePauw University 

Greencastle, Indiana 46135 

Introduction 

The use of rotenone in the reclamation of fish populations first came into use 
in the United States in 1934 (Krumholz 1948). Since then, the eradication of 
undesirable fishes from ponds, lakes, and streams by the use of toxicants has 
become an accepted fisheries management practice. Forty-nine states, including 
Indiana, have reclaimed their waters by chemical methods, mainly since 1953 (Len- 
non et al. 1971). 

In the summer of 1966, Eagle Creek and its tributaries were surveyed prior to 
impoundment by the Division of Fish and Game of the Indiana Department of 
Natural Resources to determine the relative abundance of "undesirable" fish 
species, i.e., carp, Cyprinus carpio, and gizzard shad, Dorosoma cepedianum. It was 
concluded from the study that, "no part of the watershed of Eagle Creek Reservoir 
could be omitted from an eradication project" (McGinty 1966). 

Twelve years later, Eagle Creek was chosen as an experimental study stream 
in a Model Implementation Program (MIP) initiated by the U.S. Environmental 
Protection Agency and the Department of Agriculture. The goal of the study was 
to assess the effects of improved agricultural land-use practices on the water quali- 
ty and stream biota of Eagle Creek. 

A comparison of the data collected prior to the reclamation and impoundment 
of Eagle Creek with that gathered following yields some disturbing results and 
raises some serious questions about the validity of this accepted and commonly 
used reservoir management practice. 

Methods and Materials 

Fish were collected from 16 stations on Eagle Creek and its tributaries by the 
Division of Fish and Game of the Indiana Department of Natural Resources from 8 
to 17 August 1966. Ten stations were located in the stream proper, whereas the re- 
maining six consisted of adjacent gravel pits and a farm pond (Figure 1). Rotenone 
was used as a toxicant. 

During the summers of 1978, 1979, and 1980, nine stations on Eagle Creek and 
its tributaries were sampled at least twice each summer. Four other sites were 
sampled only once during the three-year period (Figure 1). Fish were collected 
using a 30-foot, A.C. electroseine, a D.C. electric boat-shocking apparatus, and a 
one-eighth-inch-mesh nylon seine. Most of the fish collected were identified, 
weighed, and measured in the field and returned to the stream. Voucher specimens 
for many of the species are kept in the museum at DePauw University. Forty-nine 

208 



Ecology 



209 




FIGURE 1. Map of Eagle Creek drainage showing 1966, and 1978-1980 sampling 
stations. Closed circles (9) denote 1966 stream stations, closed triangles (A) 
denote 1966 gravel pits and/or a farm pond, open circles (O) denote 1978-1980 

stream stations, and half-closed circles (€)) denote 1966 and 1978-1980 stations 

sampled alike. 



210 Indiana Academy of Science 

individual collections were made during the three-year period. Common and scien- 
tific names of fishes follow Bailey et al. (1970). 

Results 

Data from the 1966 survey were obtained from an Indiana Department of 
Natural Resources report that, with one exception, listed only the species collected 
at each station (McGinty 1966). Therefore, to compare those data with the 
1978-1980 data, values are expressed in percentage frequency of occurance at 
sampling stations, that is, the number of stations at which a species was collected 
divided by the total number of stations collected, times 100. Six stations from the 
1966 survey were excluded from the calculations (i.e., adjacent gravel pits and a 
farm pond) because comparable collections were not made in the 1978-1980 study. 

All stream stations sampled in the 1978-1980 study were included in the 
calculations. Data from four stations located close to one another on the mainstem 
of Eagle Creek, and sampled only once in 1978, were grouped together as one col- 
lection. 

Of the 49 species taken during both surveys, 43 were taken in the 1966 study, 
whereas 37 were collected in the 1978-1980 survey (Table 1). Five species: gizzard 
shad, rosy-face shiner Notropis rubellus, quillback Carpiodes cyprinus, spotted 
sucker Minytrema melanops, and black crappie Pomoxis nigromaculatus were col- 
lected only from gravel pits and a farm pond during the 1966 survey. Noteworthy 
changes in the percentage frequency of occurance following the treatment and 
impoundment include redfin shiner Notropis umbratilis (from 10 to 80%) and 
bluegill Lepomis macrochirus (from 40 to 90%). 



Table 1. A combined list of species collected from the Eagle Creek drainage 
during 1966 and 1978-1980 surveys. 



Common name 


Scientific name 


1966 


1978 


1979 


1980 


Herrings 


Clupeidae 










Gizzard shad 


Dorosoma cepedianum 


X 


X 




X 


Pikes 


Esocidae 










Grass Pickerel 


Esox americanus vermiculatus 


X 


X 


X 


X 


Minnows and carp 


Cyprinidae 










Stoneroller 


Campostoma anomalum 


X 


X 


X 


X 


Carp 


Cyprinus carpio 


X 


X 


X 


X 


Silverjaw minnow 


Ericymba buccata 


X 


X 


X 


X 


Hornyhead chub 


Nocomis biguttatus 


X 








Golden shiner 


Notemigonus crysoleucas 


X 








Emerald shiner 


Notropis atherinoides 


X 








Striped shiner 


Notropis chrysocephalus 




X 


X 


X 


Common shiner 


Notropis cornutus 


X 








Rosyface shiner 


Notropis rubellus 


X 








Spotfin shiner 


Notropis spilopterus 


X 




X 




Sand shiner 


Notropis stramineus 


X 








Redfin shiner 


Notropis umbratilis 


X 


X 


X 


X 


Suckermouth minnow 


Phenacobius mirabilis 


X 






X 


Southern redbelly dace 


Phoxinus erythrogaster 


X 






X 


Bluntnose minnow 


Pimephales notatus 


X 


X 


X 


X 


Fathead minnow 


Pimephales promelas 




X 






Blacknose dace 


Rhinichthys atratulus 


X 








Creek chub 


Semotilus atromaculatus 


X 


X 


X 


X 



Ecology 



211 



Table 1. Continued. 



Common name 


Scientific name 


1966 


1978 


1979 


1980 


Suckers 


Catostomidae 










Quillback 


Carpiodes cyprinus 


X 








White Sucker 


Catostomous commersoni 


X 


X 


X 


X 


Creek chubsucker 


Erimyzon oblongus 


X 


X 


X 


X 


Northern hog sucker 


Hypentelium nigricans 


X 








Spotted sucker 


Minytrema melanops 


X 


X 


X 


X 


Golden redhorse 


Moxostoma erythrurum 


X 








Catfishes 


Ictaluridae 










Black bullhead 


Ictalurus me las 


X 


X 


X 


X 


Yellow bullhead 


Ictalurus natalis 


X 


X 


X 


X 


Brindled madtom 


Noturus miurus 


X 








Killifishes 


Cyprinodontidae 










Blackstripe topminnow 


Fundulus notatus 


X 






X 


Silversides 


Athermidae 










Brook silversides 


Labidesthes sicculus 


X 






X 


Bass and Sunfishes 


Centrarchidae 










Rock bass 


Ambloplites rupestris 


X 


X 


X 


X 


Green sunfish 


Lepomis cyanellus 


X 


X 


X 


X 


Bluegill 


Lepomis macrochirus 


X 


X 


X 


X 


Longear sunfish 


Lepomis megalotis 


X 


X 


X 


X 


Redear sunfish 


Lepomis microlophus 




X 






Smallmouth bass 


Micropterus dolomieui 


X 




X 


X 


Spotted bass 


Micropterus punctulatus 




X 


X 


X 


Largemouth bass 


Micropterus salmoides 


X 


X 


X 


X 


White crappie 


Pomoxis annularis 


X 


X 


X 




Black crappie 


Pomoxis nigromaculatus 


X 






X 


Perches 


Percidae 










Greenside darter 


Etheostoma blennioides 


X 






X 


Rainbow darter 


Etheostoma caeruleum 


X 


X 


X 


X 


Fantail darter 


Etheostoma flabellare 


X 








Johnny darter 


Etheostoma nigrum 


X 


X 


X 


X 


Orangethroat darter 


Etheostoma spectabile 




X 


X 


X 


Logperch 


Percina caprodes 


X 


X 


X 


X 


Blackside darter 


Percina maculata 


X 


X 


X 


X 


Sculpins 


Cottidae 










Mottled sculpin 


Cottus bairdi 




X 


X 


X 


Total number of species 




43 


29 


28 


33 



Notable Additions 

Of those six species collected during the 1978-1980 study, but not found in the 
1966 survey, four deserve further explanation. 

Striped shiners Notropis chrysocephalus were taken at 90% of the 1978-1980 
stations. It was probably identified as Notropis comutus the common shiner in the 
1966 study, as it occured at all of the stations surveyed. Gerking (1945) recognized 
N. chryosocephalus only as a subspecies of N. comutus (N. comutus 
chrysocephalus) in his work. 

Spotted bass Micropterus punctulatus were collected at 90% of the post-treat- 
ment stations. This species was originally identified in Eagle Creek by Gerking 
(1945). It may have possibly been overlooked in the 1966 survey. 

Orangethroat darters Etheostoma spectabile were collected at all of the 



212 Indiana Academy of Science 

1978-1980 stations. It was probably confused with the closely related rainbow 
darter E. caeruleum in the 1966 study. 

Mottled sculpins Cottus bairdi, collected at 70% of the 1978-1980 survey sta- 
tions, were notably absent from the 1966 survey. 

The remaining two species additions, fathead minnow Pimephales promelas 
and redear sunfish Lepomis microlophus, were present at only 10% of the stations. 
Their presence is probably due to introduction as bait species by fishermen for the 
former, and as a stocked species in the latter case. 

Notable Deletions 

Perhaps of greater significance are the species taken in 1966 that were not 
found in the 1978-1980 survey (Table 1). It should be noted here that failure to find 
those species in 1978-1980 does not necessarily verify their extirpation, it means 
only that they were taken in one series of samples and not in another. What is im- 
portant is that those species, some of which comprised a major portion of the 
systems biomass, have been either greatly reduced in numbers or extirpated. The 
result is a significant change in the fish community of Eagle Creek. 

Northern hog suckers Hypentilium nigricans were captured at 80% of the 
1966 stations. Their absence is particularly noteworthy since Eagle Creek appears 
to have an ideal habitat (e.g., deep riffles, rocky bottoms, and permanent flow) for 
this species. 

Golden redhorse Moxostoma erythrurum were taken from 70% of the 1966 
stations. Again, the deep pools, permanent rocky substrate, and constant flow cur- 
rently found in Eagle Creek would appear to favor the presence of this species. 

Fantail darters Etheostoma flabellare were captured at 50% of the 1966 sites. 
This species was not taken at any of the 1978-1980 stations or any of the 1966 sta- 
tions sampled in the 1978-1980 survey. 

With the exception of the common shiner discussed earlier, none of the seven 
species of minnows missing from the 1978-1980 list were taken from more than 20% 
of the 1966 stations. Nevertheless, their absence is significant since they are com- 
mon components of streams found in this area. The absence oiNoturus miurus, the 
brindled madtom, is also noteworthy as it was listed as the fifteenth most abundant 
species by weight in an "average stream environment" of Eagle Creek. 

To illustrate changing patterns in species abundance further, weight data 
from a 1966 station on the mainstem of Eagle Creek that was considered to "typify 
average stream environment for this area," was compared with a series of collec- 
tions made in this same area and upstream during 1978 (Table 2). Although sampl- 
ing techniques and exact locations of the collections differ, the results probably 
reflect a trend in species composition and abundance. 

Of the top four species collected in 1966, the number one species, golden 
redhorse, and the number four species, northern hog sucker have not been cap- 
tured to date. Furthermore, only three specimens of the second most abundant 
species by weight, smallmouth bass Micropterus dolomieui have been collected, 
two of which were young-of-the-year. Smallmouth bass were taken from 50% of the 
sampling stations in the pretreatment survey. 

In the 1978 collections, carp, one of the undesirable target species, ranked 
number one in biomass abundance, whereas the other target species, gizzard shad, 
was number eight. That trend, no doubt, is influenced by the reservoir. The 



Ecology 213 

number two species white sucker Catostomous commersoni, found at 70% of the 
1966 stations, was taken from all of the 1978-1980 sites. 

Discussion 

The reclamation of streams in the United States through the use of toxicants 
has enjoyed only limited success. Numerous authors have noted that complete fish 
kills in streams rarely, if ever, occur and that reinfestation by rough, or "undesir- 
able" fish is rapid (cf. Lennon, et al. 1971). 

Data gathered in this and other related studies support these findings. Two 
surveys of the Eagle Creek reservoir by the Indiana Department of Natural 
Resources Fish and Game Division in 1975 listed gizzard shad as the most abundant 
species captured both by number and by weight, whereas carp ranked sixth in 
numerical abundance and second in biomass abundance. A later 1978 IDNR survey 
of the reservoir showed that the gizzard shad was third by number and first by 
weight, while the carp ranked fifth by number and first by weight. The 1978 data 
gathered for this study (Table 2) seem to support the trend. Some rough fish 
species (i.e., carp) are relatively resistant to toxicants and this resistance, coupled 
with their extremely high fecundity rates, allows them to increase rapidly both in 
size and in numbers. Therefore, any reduction of rough fish is made up often in the 
first breeding season following the eradication (Hubbs 1963). 

More recently, some serious questions have been raised about the validity of 
using poisons for the control of non-game fish species (Hubbs 1963, Becker 1975, Li 
1975, Pister 1976, and Moyle 1978). The general assumption underlying nongame 
fish control, that game fish populations or growth rates are being limited by competi- 
tion and/or predation from nongame fish, is at best questionable. Moyle (1978) 
reported a growing body of evidence demonstrating that most game fish can hold 
their own, and may serve to check nongame fish populations under "natural condi- 
tions," that is, in the absence of excessive harvest or habitat alteration. Further- 
more, the contention that competition and predation between nongame fish and 
game fish is severely limiting game fish production is based largely on inferences 
from limited quantitative and qualitative data (Moyle 1977a, 1977b). 

Table 2. A relative comparison of the ten most abundant species by weight 
collected from a typical stream environment of Eagle Creek on 9 August 1966 and 

15 September 1978. 

1966 1978 

Species Weight (kg) Species Weight (kg) 

Golden redhorse 11.22 Carp 7.04 

Smailmouth bass 2.95 White sucker 3.98 

Common shiner 2.40 Longear sunfish 2.49 

Northern hog sucker 1.69 Rock bass 1.82 

Yellow bullhead 1.15 Spotted sucker 1.10 

Rock bass 0.76 White crappie 0.84 

Largemouth bass 0.76 Creek chub 0.81 

Longear sunfish 0.61 Gizzard shad 0.26 

White crappie 0.37 Grass pickerel 0.24 

Stoneroller 0.33 Yellow bullhead 0.20 



214 Indiana Academy of Science 

That game fish may actually benefit from a commensal feeding relationship 
with rough fish is implied by Reighard (1920) and Greely (1935). Both reported that 
smallmouth bass take a position downstream from hog suckers and feed on drifting 
aquatic insects disloged as the hog sucker turns over rocks. Larimore et al. (1952) 
noted a strong correlation (r = 0.85) between the weight distribution of small- 
mouth bass and hog suckers. Perhaps the current paucity of smallmouth bass in 
Eagle Creek is due, in part, to the absence of hog suckers from the system. 

In general, the interactions between game fish and non-game fish are poorly 
understood. A need exists for sound ecological analysis of these relationships as a 
basis for thoughtful and sound fisheries management programs. Pister (1976) con- 
cluded that, "'Management' has been manifested in nongame species destruction, 
often with virtually no biological justification, . . .(while) little effort has been 
expended by agency fishery biologists in researching and understanding inter- 
specific relationships." 

The present strategy of reservoir management wherein the fish community is 
disrupted by rotenone for the control of a few species is questionable. The present 
reservoir preimpoundment eradication program should be re-evaluated. Less 
destructive reservoir management techniques exist for the control of rough fish 
(i.e., construction of barrier dams and the institution of maximum size limits on 
game fish, etc.) and may provide additional solutions. 

The recent passage of the Forsythe-Chafee nongame fish and wildlife conser- 
vation act (PL 96-366) can provide the needed impetus for the careful conservation 
and management of our nongame resources. Pister (1976) has stressed the need for 
the management of nongame fish and wildlife, primarily in response to the pro- 
jected increasing demand by the populace for outdoor oriented recreation. Thus, 
the fisheries biologist must be concerned with a broader portion of the aquatic 
biota. Moyle (1978) recognized this need for the management of all fish," ". . . 
fisheries biologists should start considering themselves as stewards of ecosystems 
and native faunas, as well as managers of game fish populations for consumption." 

Acknowledgments 

We are extremely grateful to Dr. Louis A. Krumholz and Dr. William D. Pear- 
son for their reviews of the manuscript. We would also like to thank all those peo- 
ple who assisted in the field work and the compilation of the data. This study was 
supported by a grant from the U. S. Environmental Protection Agency (Project no. 
R-806-164). 

Literature Cited 

1. Bailey, R. M., J. E. Fitch, E. S. Herald, E. A. Lachner, C. C. Lindsey, C. R. 
Robins, and W. B. Scott. 1970. A list of common and scientific names of fishes 
from the United States and Canada. 3rd Ed. Amer. Fish. Soc. Spec. Publ. No. 
6:1-150. 

2. Becker, G. 1975. Fish toxification: biological sanity or insanity? pp. 41-53. In 
P. H. Eschmeyer (ed.), Rehabilitation of fish populations with toxicants: a sym- 
posium. N. Cent. Div. Amer. Fish. Soc. Spec. Publ. 4. 

3. Gerking, S. D. 1945. The distribution of fishes in Indiana. Invest. Indiana 
Lakes & Streams. 3(1): 1-137. 

4. Greely, J. R. 1935. Fishes of the watershead. In A biological survey of the 
Mohawk Hudson watershed. Suppl. 24th Ann. Rept. N. Y. Consv. Dept. 
1934(1935): 63-101. 

5. Hubbs, C. 1963. An evaluation of the use of rotenone as a means of 
"improving" sports fishing in the Concho River, Texas. Copeia. 1973(1): 
199-203. 



Ecology 215 

6. Krumholz, L. A. 1948. The use of rotenone in fisheries research. J. Wildl. 
Mgmt. 12(3): 305-317. 

7. Larimore, R. W., Q. H. Pickering, and L. Durham. 1952. An inventory of the 
fishes of Jordon Creek, Vermillion County, Illinois. 111. Nat. Hist. Surv. Biol. 
Notes No. 29: 1-26. 

8. Lennon, R. E., J. B. Hunn, R. A. Schnick, and R. M. Burress. 1971. Reclama- 
tion of ponds, lakes, and streams with fish toxicants: a review. FAO, U. N. 
Fish. Tech. Pap. 100. 

9. Li, H.W. 1975. Competition and coexistence in stream fish, pp. 19-30, In P.B. 
Moyle and D.W. Koch, (eds.), Symposium on trout/nongame fish relationships 
in streams Des. Res. Inst. Misc. Rept. No. 17. 

10. McGinty, D. J. 1966. Pre-impoundment fish population study of the Eagle 
Creek reservoir watershed. Indiana Dept. Nat. Res. Div. Fish & Game. 
Mimeo. 15pp. 

11. Moyle, P. B. 1977a. In defense of sculpins. Fisheries. 2(1): 20-23. 

. 1977b. Are coarse fish a curse. Fly Fisherman. 8(5): 35-39. 

. 1978. Evaluation of nongame fish poisoning operations. Calif. Dept. 

Fish & Game Rept. Mimeo. 17pp. 

12. Pister, E. P. 1976. A rationale for the management of nongame fish and 
wildlife. Fisheries. 1(1): 11-14. 

13. Reighard, J. 1920. The breeding behavior of the suckers and minnows. Biol. 
Bull. 38(1): 1-32. 



Alopecurus pratensis L. Discovered in Porter County 

C. L. Rhykerd, E. L. Hood, B. 0. Blair, Ekpo Ossom, 
and J. W. Lightner 

Department of Agronomy, Purdue University 
West Lafayette, Indiana 47907 

Introduction 

Alopecurus pratensis L., meadow foxtail, is a long-lived perennial grass which 
is native to the temperate portions of Europe and Asia. It was first cultivated in 
Europe about 1750 and is still grown as a forage crop on poorly drained soils of high 
fertility. Meadow foxtail is believed to have been introduced into northeastern 
United States and eastern Canada in the latter part of the nineteenth century. It 
has been grown to only a very limited extent in the northcentral and northeastern 
states although it has recently become of some importance in the Pacific North- 
west. To the authors' knowledge, the only report of meadow foxtail being grown in 
Indiana is that reported in Deam's book "Grasses of Indiana" (1). According to 
Deam, meadow foxtail has been reported growing in Tippecanoe County which was 
most likely a seeding by a Purdue University agronomist. 

Plant Description and Closely Related Species 

Meadow foxtail is a long-lived perennial cool-season grass possessing an in- 
florescence which is very similar in appearance to that of timothy {Phleum 
pratense L.). It produces comparatively few rhizomes ranging from approximately 
five to ten cm. in length resulting in loose tufts with numerous basal leaves. 
Growth starts extremely early in the spring and it is one of the earliest grasses to 
head. The erect flowering culms generally are about one meter high but have been 
reported to occasionally reach a height of nearly two meters (2). Meadow foxtail 
seed is small, light and fluffy making it difficult to harvest as well as to sow. 
Meadow foxtail, because of its common name, should not be confused with some of 
the weedy grasses. It bears little, if any, resemblance to other weedy grasses refer- 
red to as foxtail. Meadow foxtail is a high quality forage crop with no evidence of 
weedy characteristics (3). 

Two other species of Alopecurus are found in Indiana. A. aequalis Sobol., 
short-awn foxtail, is occasionally found growing in central and northern Indiana in 
shallow water, pond borders, and swamps that become dry in midsummer. A. 
carolinianus Walt, can be found throughout most of Indiana. It is found growing on 
mucky soils and in ditches in northern Indiana while in southern Indiana it will be 
found on slightly acid, light-colored clay soils especially in fallow fields (1). 

Adaptation 

Meadow foxtail is adapted primarily to a cool and moist climate. However, it 
can be grown in regions where summer temperatures occasionally reach 100° F as 
well as regions where winter temperatures drop below 0° F for long periods. Piper 
(2) reported that "it is perhaps the most winter hardy of any cultivated perennial 
grass." In addition, it withstands cold weather in early spring after its growth has 
begun better than any other grass. Consequently, it survives well where frost may 
occur any month of the year as at high elevations or muck areas in northern United 
States where frost can occur any month of the year. 

216 



Ecology 217 

The best growth of meadow foxtail is obtained on fertile, moist, or swampy 
soils. Muck or peat soils that are swampy or are overflow lands are ideal for 
meadow foxtail. It grows well under irrigation but does not withstand drought. 

Utilization 

Meadow foxtail is primarily a pasture grass in northern regions where it is 
grown (3). Cattle and sheep graze it more readily than most other grasses due to its 
palatability and high percentage of basal leaves. This can be a disadvantage when 
grown in a mixture with other grasses of lower palatability as livestock will graze 
it very closely allowing the less palatable species to predominate. 

Due to its low yield and high percentage of basal leaves, meadow foxtail is 
seldom used for hay. However, in northern Europe it is a favorite hay crop on wet 
meadows. Although yields of only one to three tons are most common, yields as 
high as 4-Vz tons per acre have been reported in England and France (2). 

Several Ecotypes Discovered in Porter County 

In the spring of 1975, a few scattered plants of meadow foxtail were observed 
on the muck weather station of the Pinney-Purdue Agricultural Center at 
Wanatah, Indiana in Porter County. At first the plants were assumed to be a very 
early selection of timothy, but the plants later were identified as meadow foxtail. 
The site has been clipped only once or twice annually over the past 5 years in order 
to observe the meadow foxtail plants. Meadow foxtail appears to be very com- 
petitive with Kentucky bluegrass under these conditions and now occupies about 
20 percent of the area in the muck weather station. Another closely related but 
much shorter species, either A. aequalis Sobol. or A. carolinianus Walt, also has 
been observed growing on the Pinney-Purdue Agricultural Center as a weed in 
cultivated fields. 

It is believed that this meadow foxtail originated as a contaminant of a forage 
grass seeded in an evaluation study adjacent to the muck weather station in the 
late 1960s. Several ecotypes have been transplanted to the Agronomy Farm, West 
Lafayette, to observe their growth characteristics as compared to that of timothy. 
Considerable differences have been observed among the meadow foxtail ecotypes 
relative to leaf width and density of basal leaves. These ecotypes of meadow foxtail 
head out in late April to early May, being three to four weeks earlier than timothy. 

Potential for Meadow Foxtail 

Meadow foxtail is almost unknown to Corn Belt forage researchers as well as 
forage producers. This is probably due to its historically low yielding ability as well 
as its lack of persistence under droughty conditions. However, some of the 
characteristics observed from the ecotypes growing in Porter County would 
indicate that meadow foxtail could be a valuable forage species on poorly drained 
soils in northern and possibly central Indiana. 

Many farmers on the nearly one million acres of muck soils in northern 
Indiana have given up trying to grow corn and/or soybeans. This is due to the 
problems encountered in getting the crops planted in the spring because of ex- 
cessive moisture along with the often short growing season resulting from early 
frosts on these low-lying soils. Some farmers are presently using these soil for 
pasture. Kentucky bluegrass iPoa pratensis L.) is very well adapted to these soils 
as is birdsfoot trefoil (Lotus corniculatus L.), a legume. However, Kentucky 
bluegrass is extremely competitive and, if not properly managed, will out-compete 
the birdsfoot trefoil and necessitate the application of nitrogen fertilizer to obtain 



218 Indiana Academy of Science 

high yields. Since meadow foxtail appears to be well adapted to these soils but 
less competitive than Kentucky bluegrass, it may be an ideal grass to grow in 
association with birdsfoot trefoil. The very early spring and late fall growth of 
meadow foxtail along with the high late spring and summer production of birds- 
foot trefoil should provide a highly palatable pasture over a long grazing season. 

In addition, meadow foxtail may be a desirable grass for northern Indiana 
dairy farmers to seed with alfalfa. A limited amount of grass in a mixture with 
alfalfa can have many benefits, such as better erosion control and easier 
harvesting and drying, when compared to pure alfalfa without reducing the quali- 
ty of the hay or silage. 

Summary 

The observations reported in this paper can be summarized as follows: 

1. Several ecotypes of meadow foxtail have been discovered growing on the 
shallow muck soils in Porter County. 

2. Meadow foxtail closely resembles timothy in the heading stage. 

3. In northern Indiana, meadow foxtail heads out nearly one month earlier than 
timothy. 

4. These ecotypes of meadow foxtail are spreading in competition with Ken- 
tucky bluegrass. 

5. Meadow foxtail appears to be well adapted to the muck soils of northern 
Indiana where frosts can occur any month of the year. 

6. Meadow foxtail may have promise in northern and possibly central Indiana 
as a grass to grow in association with 

A. birdsfoot trefoil for pasture particularly on muck soils. 

B. alfalfa for hay or low moisture silage by dairymen. 

Literature Cited 

1. Deam, C. C. 1929. Grasses of Indiana. Publication No. 82. Division of 
Forestry, Department of Conservation, State of Indiana. 

2. Piper, C. V. 1924. Forage Plants and Their Culture. Rev. Ed. The MacMillan 
Co., New York. 

3. Wheeler, W. A. 1950. Forage and Pasture Crops. Van Nostrand, New York. 



ENGINEERING 

Chairman: Donald D. Gray 
School of Civil Engineering, Purdue University, West Lafayette, Indiana 47907 

Chairman-Elect: Mark H. Houck 
School of Civil Engineering, Purdue University, West Lafayette, Indiana 47907 

ABSTRACTS 

Development of a Robust Routine for Calculating Vapor-Liquid Equilibrium Con- 
ditions. Warren W. Bowden, Department of Chemical Engineering, Rose-Hulman 

Institute of Technology, Terre Haute, Indiana 47803. Reliable routines for the 

calculation of vapor-liquid equilibrium conditions are a necessity for the design of 
equipment for such operations as flash separation, distillation and absorption. This 
paper describes an efficient and robust algorithm for this type of calculation. 

Here 'efficient' means that the number of iterations is not large and 'robust' 
means that convergence to an unambiguous solution occurs for appropriate star- 
ting values. 

The algorithm involves calculation of K-factors using the Lee-Kesler equation 
of state. Composition effects are introduced by the methods of Plocker, Knapp and 
Prausnitz (1978). The equation of state is applied twice, once to the liquid phase and 
again to the vapor phase, to evaluate the fugacity of each component in both 
phases. This calculation requires an iterative solution of the equation of state to 
determine the liquid- and vapor- densities. Hence, the calculation involves a double 
iteration: an outer loop where temperature (for example) is the variable and an 
inner loop when density is the variable. At moderate conditions the calculation pro- 
ceeds without difficulty. At more severe conditions, especially as the critical point 
of the mixture is approached, the calculation quite commonly fails. A robust 
routine must have provisions for appropriate starting temperature- and density- 
values and carefully constructed iterative procedures. 

This paper describes a) the features of the algorithm which make it robust: 
choice of initial values, iteration techniques, and b) some results obtained with the 
program. 

Combined Sewer Separation: Case Study. Christopher B. Burke, School of Civil 
Engineering, Purdue University, West Lafayette, Indiana 47907. After Con- 
gress enacted the Water Pollution Control Act (PL 72-500), much of the nation has 
been required to eliminate combined sewer overflows. Different approaches have 
been proposed and implemented. Three of the approaches are: 1) the upgrading or 
design of a sewage treatment plant or interceptor to handle the combined sewage 
during storms 2) temporarily store the combined sewage or 3) physically separate 
the stormwater from the sanitary wastes. This paper will take a look at the third 
alternative as it was applied to an urban area. 

The project was undertaken for Lansing, Illinois a suburb located southeast of 
Chicago. The engineer was R. W. Robinson and Associates in South Holland, 
Illinois. The storm sewer system and detention pond were designed using the Il- 
linois Urban Area Simulator (ILLUDAS). The total drainage area separated was 
about 460 acres. 

219 



220 Indiana Academy of Science 

Microcomputer Driven Multi-Point Controller. David D. Chesak, Department of 

Physics, St. Joseph's College, Rensselaer, Indiana 47978. There has been a 

marked increase in the use of microcomputers in industrial and academic settings. 
While many of these are used as programmable controllers, it is felt that further 
advances can be made. The interface circuit described in this paper is intended to 
provide a simple means for digital control of multi-element devices such as pipe 
organs, incandescent light displays or control valves in a robot. 

The interface circuit requires only one eight bit port from the computer to 
drive hundreds of elements in an on-off, two state mode. The switching operations 
are accomplished in microseconds. Any computer that provides an eight bit port 
can be programmed in machine code or any of the more sophisticated languages to 
drive the interface. Machine code provides the fastest switching speeds while a 
language such as BASIC provides greater ease of programming. 

The circuit described has been used to play a three octave electronic organ. 
The program in BASIC compiles the note sequencing from a data code taken from 
sheet music. There is no particular limitation on the number of notes that can be 
sounded simultaneously. The microcomputer uses the very flexible S-100 bus 
system with an 8080 CPU. As many as 96 notes can be controlled with the circuit 
described and, with a minor modification, up to 768 devices can be controlled. The 
integrated circuits are the TTL type and all are available from a number of 
manufacturers. 

Chemical and Physical Characterization of Wastewater Sludges in Indiana. Wayne 

F. Echelberger, Jr. and J. C. Randolph, School of Public and Environmental 
Affairs, Indiana University, Bloomington, Indiana 47405, J. Michael Jeter, Indi- 
anapolis Water Company, and Steve W. Kim, Division of Water Pollution Control, 

Indiana State Board of Health, Indianapolis, Indiana 46206. This research 

study was directed at obtaining an accurate and as comprehensive as possible 
inventory of the quantity and quality of municipal and industrial wastewater treat- 
ment sludges being generated throughout the State of Indiana. The specific pro- 
cedures for data gathering, analysis and computerization were developed and ex- 
ecuted through close and continuing coordination with the Indiana State Board of 
Health (ISBH). 

In order to characterize the sludges generated as a result of the treatment of 
municipal wastewater in the state, sample sites were selected on the basis of 
population, with the sludges being collected and analyzed for both compatible and 
incompatible pollutants. The data presented in this survey indicates in detail the 
chemical concentrations in these residuals from wastewater facilities serving 90 
percent of the sewered population in Indiana. 

Overall, the results of this investigation indicate that municipal sludge is quite 
variable in terms of nitrogen, heavy metals and polychlorinated biphenyl (PCB), 
with the range of concentrations commonly covering three orders of magnitude. 
Considerations pertaining to these constituents include such things as the in- 
dustrialization of the surrounding community, efficiency of the treatment pro- 
cesses, solids loading on the treatment plant and sludge handling techniques. 

A closer scrutiny of this information indicates that the majority of sites sampl- 
ed and analyzed were generally found to have relatively low concentrations of in- 
compatible contaminants. A few of the municipalities, receiving heavy industrial 
loadings from either a single source or a group of manufacturing entities, tend to 
distort the means and make these values unrealistically high. As such, the overall 



Engineering 221 

mean values are somewhat limited in their use for defining the quality 
characteristics of Indiana municipal sludges. 

For the analysis of the quality characteristics of industrial sludges resulting 
from the treatment of process wastewater, manufacturing sites were selected 
primarily on the basis of SIC codes. This was done in an attempt to draw general 
conclusions aligning manufacturing types with the presence of certain waste con- 
stituents. The sampling of these sites posed problems in terms of the variable 
nature of the wastes treated in each industry and the discretionary respone that 
was encountered from some of the companies. Ultimately, forty-nine samples were 
received and analyzed for nitrogen, solids, heavy metal and PCB concentrations. 

The statistical means, medians, and ranges for the industrial sludge analyses 
that were determined from this survey, in general, reveal tremendous variations in 
the quality characteristics for these sludges. This is no doubt due to the selective 
nature of the wastewater treatment procedures and the specific characteristic of 
the source for these constituents. The removal of plating wastes, as an example, 
will be significantly influenced by the chemicals used for the particular electro- 
plating process and the presence of other constituents will generally be a negligible 
factor. As such, the sludge quality characteristics should be regarded as unique to 
the industrial source and somewhat representative of the manufacturing process. 
Comparisons to other types of industrial activities can be done only in a very 
general sense. 

This research was jointly sponsored by the Indiana State Board of Health, U. 
S. Environmental Protection Agency, and Indiana University. 

Particulate Sedimentation in Shallow Lakes. Aldo Giorgini and Mark Smith, 
School of Civil Engineering, Purdue University, West Lafayette, Indiana. 47907. 

Particles are released from the surface of a shallow lake with idealized 

bathymetry and their motion is followed until they deposit on the lake bottom. 
Since an exact solution of the wind driven circulation in such lakes was found by 
the senior author, it has been considered possible to interpret numerically the tra- 
jectories of the particles which are usually strongly effected by numerical approx- 
imations. 

It is found that the bottom distribution of particles, as generated by a uniform 
distribution of "lighter" particles on the surface, is highly irregular and has regions 
where particles never fall. 

[Particles are defined as being "light" when their free fall velocity in the water 
is smaller than the largest upward velocity component in the lake]. 

The Boundary Conditions of Oxygen Transfer in Water with Respect to 

Temperature. Robert H. L. Howe, West Lafayette, Indiana, 47906. The three 

boundary conditions of oxygen transfer in water with respect to temperature are 
discussed: (1) when oxygen supply is provided by an excess air flow rate in a small 
volume of water under a back pressure; (2) when the air flow rate in an appreciable 
volume of water and a steady state is maintained; and (3) when the air flow rate is 
relatively small and it takes quite a long time to reach the dissolved oxygen satura- 
tion level. Mathematically, the three conditions are: (1) dc/dt = - ; (2) dc/dt = 0; 
and (3) dc/dt - + ; where C = dissolved oxygen concentration in mg/liter and t = 
time. 

The Determination of the Rising Velocity of a Gas Bubble Through a Liquid Col- 
umn by Howe's Method. Robert H. L. Howe. West Lafayette, Indiana 47906 and 



222 Indiana Academy of Science 

Hakki Dingil, Istanbul Technical University, Turkey. A mathematical deriva- 
tion for the determination of the rising velocity of a gas bubble is presented. Ex- 
amples of the experimentation and some results are discussed in comparison with 
the calculation by the Newitt-Dombrowski-Knelman Formulation. 

The equation derived by Howe can be used also to determine the fluid viscosity 
when the gas diameter and its rising velocity are known. 

Difference Models of River Reaches. P. G. Katz and G. H. Toebes, School of Civil 
Engineering, Purdue University, West Lafayette, Indiana 47907. River flow- 
watershed runoff models of the Green River Basin of Kentucky are required for ef- 
ficient operation of reservoirs in the basin. Correlative linear models that relate 
reach outflow to reach inflow and precipitation offer a relatively simple, easy to use 
method of satisfying the requirements. 

The forecasting errors of previously constructed linear models for the Green 
River basin were examined. This provided a guide to the construction of new 
models. These models relate the changes in inflows to the changes in the outflow. 
Such models, called difference models, were constructed for the 9 reaches in the 
basin. Their construction involved taking the first difference of each of the outflow 
series and relating them statistically to the first differences of the inflow series. 
Precipitation was not differenced. The submodels and the basin model made by 
linking the submodels together were found to be better than the linear models con- 
structed earlier. 

Attempts were made to improve the difference models by the addition of an 
error model and the use of multilevel difference models. None of these variants 
were found to significantly improve the accuracy of the model. 

Premultiplication of the precipitation by the average monthly runoff coeffi- 
cient was found to improve the forcasting accuracy of a difference model built on 
that basis. The use of the SCS Direct Runoff Method to estimate effective 
precipitation for input into the difference model was found to decrease the ac- 
curacy of the model. 

Analysis of Suspended Particulate Data from Chicago. A. Ramachandra Rao, 
School of Civil Engineering, Purdue University, West Lafayette, Indiana 47907. 

The air-suspended particulate data from Chicago are seasonal and nonsta- 

tionary. The nonstationary in the data is made more pronounced by regulations 
which banned the use of high sulphur coal in Chicago. 

The present study deals with modeling air-suspended particulate data from 
Chicago. A stochastic modeling technique called the 'Intervention Analysis' is used 
to develop models for these data. The modeling technique and its implications on 
policy are discussed in detail. 

Performance of the Purdue Hydromechanics Laboratory Closed Circuit Wind Tun- 
nel. Gerard F. Sheldon and Donald D. Gray, School of Civil Engineering, Purdue 

University, West Lafayette, Indiana 47907. The Hydromechanics Laboratory 

of the Purdue University School of Civil Engineering houses a large, single return, 
closed circuit, subsonic wind tunnel having two test sections in series. The tunnel is 
powered by a 150 h.p. electric motor which produces continuously variable mean 
velocities of up to 14 m/s in the large test section (3.05m long x 2.44m wide x 1.83m 
high) and up to 58 m/s in the small test section (4.57m long x 1.34m wide x 0.95m 
high). Each test section contains a remote controlled motorized traversing 
mechanism. 



Engineering 223 

Following structural modifications intended to reduce tunnel wall vibrations, 
pitot tubes and hot wire probes were used to measure mean velocity and tur- 
bulence intensity distributions in seven planes normal to the flow at three 
airspeeds. In each test section, variations in mean velocity and turbulence levels 
generally declined in the streamwise direction. At top speed, the mean velocity 
varied by 12.6% and the maximum turbulence intensity was 1.1% in the farthest 
downstream plane of the large test section (outside of the wall layers). At top speed 
in the farthest downstream plane of the small test section, the corresponding 
values were 2.8% and 0.3%. The superior performance of the small test section is 
attributable to the greater contraction ratio. The modifications were found to have 
reduced turbulence intensities by up to 86%. 



Grain Reserve Systems: Failure of Multiobjective Decisions Under 

Catastrophic Intervention, A Case for Topologic Stability in 

Singular Mapping 

Bithin Datta and Mark H. Houck 

School of Civil Engineering 

Purdue University, West Lafayette, Indiana 47907 

Introduction 

Fundamental similarities and differences exist between management of water 
reservoir and grain reservoir systems. The similarities are due to their roles as 
temporal buffer stocks for commodity fluctuations. The differences relate to the 
fact that streamflow is primarily a weather related process but grain production 
and consumption are affected by national policies, economic incentives, 
technological changes and other factors in addition to the vagaries of nature. It is 
possible to adopt stochastic or deterministic optimization, reliability analysis and 
multi-objective programming to the management of grain reserves in a manner 
similar to their use in the management of water reservoir systems. 

However, modifications are also necessary in the methods of analysis. The ma- 
jor area of emphasis in grain buffer stock policy models has been on the market 
economics, while water reservoir planning and operations models have concen- 
trated on natural phenomena with poor long term predictability. The existence of a 
market for grains is the major factor affecting the transfer of simulation and op- 
timization models to grain buffer stock management. In water resource analysis, 
streamflow is independent of the demand for water or its products, whereas the 
amount of grain produced is a function of demand for grains. Therefore, an analyst 
must model the economic aspects in particular. The grain market itself can be 
treated as implicit in historical production data series. Production then becomes a 
surrogate for technological changes as well as market and weather induced 
changes in a limited sense. 

The importance of a grain reserve is in its role as an insurance against 
catastrophes. In particular for the U. S., where major national income comes from 
grain exports, the optimum management of a grain reserve system is of prime im- 
portance. The major varieties of grains in this country which are sources of staple 
food are wheat and corn. Different grains are close substitutes on both demand and 
supply sides. It is therefore desirable that a model of the grain market should in- 
clude all the grains. Any effort to stabilize the price of one grain should affect the 
prices of other grains too. Therefore, the best method would be to include explicitly 
and simultaneously the different categories of grains in the model. Also, such a 
model should consider monthly or seasonal fluctuations and the exact location of 
reserves. However, methods and capacities of computational procedures stand in 
the way of constructing such a global model at this time. Simplifications and ap- 
proximations are adopted to cope with this problem. 

The First Model 

The foremost objective of building a grain reservoir system is to stabilize the 
price variation within the limits of profitable return to the producers. At the same 
time, the capacity of the reservoir should be offset by (i) monetary benefits to the 
farmers, (ii) price stabilization above the subsidization level, (iii) reduction in pay- 
ment to the farmers by the government through set aside program, or (iv) gains in 

224 



Engineering 225 

social welfare by assuring certain levels of consumption at all times, particularly 
for the poorer sections of the population. So the two basic objectives may be stated 
as: (1) minimize grain price fluctuations over some time horizon, and (2) minimize 
the grain reserve capacity. 

A multiple objective linear program that ensured a minimum return to 
farmers over a ten year study period (1968-77) was constructed and solved to deter- 
mine the relationships between objectives 1 and 2. The results are shown in Table 1 
where it is evident that an inverse relationship exists between the reserve 
capacities for wheat and corn and the grain price stability. For example, a wheat 
reserve capacity of 2.1 billion bushels ensures that wheat prices deviate only 
$1.86/bushel over ten years; whereas a reserve capacity of 1.95 billion bushelsj)ro- 
duces a price flucturation of $3.25/bushel. The smaller reserve capacities are not 
feasible because they do not provide the minimum return to farmers as specified in 
the model. 

Together with the above study, an effort was made to correlate the historical 
values of exports with other related variables. However, such a model exhibited 
very poor performance. It was noted that with the imposition of an export embargo 
on grains the prices of grains like corn or wheat have slumped by about 25% in a 
single day in the domestic market. This only points out the inadequacy of methods 
used in economic analysis, based on equilibrium conditions of demand and supply as 
required for models such as the linear program mentioned above. The instan- 
taneous response to external intervention in the market system certainly points to 
the divergencies and discontinuities which exist in a socio-economic system and 
which are generally ignored in the process of conventional analysis. 

The Second Model 

The above notion served as a motivation for trying to build a conceptual 
model, based on the ideas of the "Separation Theorem" as noted by Samuelson (6) 
and the unstable behavior of stock exchanges as noted by Zeeman (7). 

We start from the separation theorem in economics, which states that "Points 
of stable equilibrium (in the small) are separated by points of definitely unstable 
equilibrium and vise versa," where equilibrium "in the small" exists, if for suffi- 
ciently small perturbations the equilibrium is stable. It can also be recalled that a 
critical point is structurally stable only if it is not degenerate. [A function f has a 
nondegenerate critical point at u, if the Hessian matrix 



Table I. Possible Values of Grain Reserve Capacity vs. Maximum Price 
Fluctuation For a Ten Year Period 





Wheat 




Corn 


Capacity 


Price Fluctuation 


Capacity 


Price Fluctuation 


(million bushels) 


($/bushel) 


(million bushels) 


($/bushel) 


1900 


no feasl. sol. 


1600 


no feasl. sol. 


1950 


3.248 


1639 


2.11 


1975 


2.872 


1650 


2.07 


2000 


2.529 


1700 


1.93 


2025 


2.196 


1800 


1.78 


2050 


1.952 


1850 


1.72 


2075 


1.860 


1894 


1.674 


2100 


1.860 


1900 


1.674 


2212 


1.860 


2106 


1.674 



226 Indiana Academy of Science 

d 2 f 



Hf|. 



Hi 



is nonsingular or determinent (Hf/ ) = 0. The condition that the Hessian be non- 
singular is exactly the condition that the Jacobian of the mapping 



Df = (-JL •" il\:R n -R n 



be nonsingular, which is the condition that the graph of Df meet that of the zero 
function transversely.] 

Another mathematical property taken into account is that transversality is a 
stable property and transverse crossings are themselves stable. However, when 
dealing with a family of functions, where the paramaters are allowed to vary, a 
structurally stable family can include individual functions with degenerate critical 
points. The surrounding members of the family exert some form of calming in- 
fluence on the degenerate functions, which is called "unfolding" by Rene Thorn. As 
an example, the family of functions 

f = ^ + L. x 2 
4 2 
does not have structurally stable critical points, as the mapping (x,a) — Df(x,a) 
where Df = -?- intersects the zero plane nontransversally because the intersection 
is not a manifold. However, by adding a perturbation term bx to f, 

* x 4 x 2 u 

f + p = — + a_ + bx 

4 2 

the intersection of the mapping D(f + p) with the zero plane becomes a manifold, 
i.e. a curve in "n" dimensional space with no intersections. And the critical points of 
this family of functions as obtained by mapping in the (x, a, b) space for different 
values of the parameters a and b become structurally stable. 

The family 

x 4 x 2 u 
£_ + a_ + bx 

4 2 

is called the cusp catastrophe and the mapping (x, a, b) — Df(x, a, b) is called the 
catastrophic manifold. Singular points of this mapping are those points with ver- 
tical tangents when x is assumed as the vertical axis. Therefore, it can be concluded 
that by adding a perturbation term, the critical points of a family of functions ac- 
quire some form of topologic stability. 

Rene Thorn showed in his "Catastrophe Theory" that "Structurally stable 
smooth change of coordinates can occur in a R parameter family only in finite 
ways." These families of mappings are called catastrophic manifolds. Because 
repeatability and reproduceability are the main ideas of any modeling effort, we 
shall use the above concept together with the "Separation Theorem", trying to pre- 
sent a conceptual model. The problem becomes the exploration of equilibrium sur- 
faces which are connected to the stability changes of the system, as the control 
parameters slowly evolve. The correlation between the Catastrophe Theory and 



Engineering 227 

the Bifurcation Theory, embedded in the perturbation concept, can help remove 
the qualitative nature of the Catastrophe Theory. 

We now consider this situation of a sudden restriction in export commitments 
by government, and its effects on the price stability of the grain market. It is 
assumed that initially we have a stable system. Therefore, in the chronological 
dynamic model we considered before, the state of the system at a particular 
discrete time interval was supposed to be related to the state of the system at a 
previous time interval and other related variables. The shock to which the system 
is subjected may cause a sudden shift in the equilibrium position to another 
equilibrium position, through a short lived process of dynamic instability. 

Our main concern is to investigate these two states of equilibrium separated 
from each other, or the entire process of comparative statics. Here we confront the 
problem of establishing the form and mechanism of this intermediate path, where a 
bifurcation occurs through external perturbations. The concept of structural 
stability in Catastrophe Theory, may form a link between the static stationary 
states and truly dynamic path through time under external shocks. The whole pro- 
cess is assumed to be structurally stable, repeatable or reproduceable. The main 
objectives are to construct some sort of a predictive model, to design effective cor- 
rective measures, to make the rate of change of the price index for grain positive, 
and to remedy the catastrophic slump in the price index. 

The following variables are defined. 

Pj o price index for grains (may be for aggregated average of all varieties 
or a particular one such as wheat) 



= Pj - rate of change of price index 



T = dP 
dt 

T = — initial stable condition 
T > — export market dominates 
T < — domestic market dominates 

(Excess stock expected to be left over\ /Export commitments\ 
after domestic consumption and j-( to other I 
reserve commitments / \ countries / 

(Total domestic consumption) 
X = excess demand in domestic market 
X = — no excess demand in domestic market 

The following hypotheses based on the behavior of the economic system are 
put forward. 

Hypothesis 1: T responds to K and X much faster than K and X respond to T. This 
implies that price stability is affected much faster by a change in the proportion of 
expected net excess of grains to that of domestic consumption and the excess de- 
mand in the domestic market. 

Hypothesis 2: If K is small, T is a continuous, monotonic increasing function of X, 
passing through the origin. Therefore, when the ratio K is small, T will try to in- 
crease with a positive change in excess domestic demand. This is true for both T < 
and T > 0. 

Hypothesis 3: If K is large, this will cause some instability in the whole system. 
Thus for a large K, where net excess is large, perturbations in the form of an ex- 
ogeneous shock may cause a change in the equilibrium state of the system, i.e. a 
jump from one stable state to another. The evidence of postulation of such a sudden 



228 



Indiana Academy of Science 



crash is inherent in the "Separation Theorem." The basic ideas in the three 
hypotheses may be summarized by Figures 1 and 2. 



T 4 



crash 



Export market 

dominates) 



(Domestic market (For large K) 
dominates) 



I (For small K = 0) 

Figure 1. Synthesis of the Hypotheses 

-7. 





Exports dominate 
(T>0) 

[DV=T 3 -(K-K o )T-X=0] 



(K-X) 
Plane o 
Control Space 



\ 



Domestic mar- 
ket dominates 
(T<0) 




Bifurcation set 



Figure 2. The Behavior and Control Surfaces 



The behavior surface is represented by the function 



DV 



(K 



K )T 



X 



[K Q = some normal value of K] 



where V 



(K 



K ) T 2 



XT represents the state of the system in some form. 



The mapping of (T, K, X) — DV(T, K, X) represents the nature of flow in the behavior 
surface due to changes in the control parameters K and X. The singular points of this 
mapping are enclosed within the bifurcation set shown in the control space. The 
basic conclusion we can dervie from the above hypotheses is that in order to stabilize 
prices, measures should be adopted to decrease K and increase X when a sudden col- 
lapse is imminent. However, transition back, as shown by arrows, is a slower pro- 
cess. Also, since catastrophic conditions fortunately do not occur very often, 



Engineering 229 

calibration may be difficult to obtain. We may look upon the catastrophic manifolds 
in the simplest sense as an effective way of describing a system with variable 
degrees of hysteresis. 

Summary and Conclusions 

The entire economic system of grain production, distribution, consumption, 
storage and export is by no means a simple isolated entity. The crucial interactions 
of various social, political and economic forces are fundamental to the system. 
Therefore, an intuitive understanding of the entire process based on logical 
assumptions should yield a better model than other much more sophisticated 
analytic techniques. It is strongly believed that the above process of exploration of 
singularities of mapping and structural stability in an economic system should give 
a better idea of the interactions of crucial variables at crucial states, and should 
help to identify effective corrective measures, both quantitatively and qualitatively. 
The conceptual model based on the ideas of Catastrophe Theory was presented as a 
supplement to the previously described multi-objective optimization model, which 
fails to operate meaningfully under catastrophic conditions marked by sudden 
changes in the stable configuration of the system. The assumptions involved are no 
doubt partly intuitive and based on some forms of topologic indefiniteness. Also, it 
is plausible to find limitations of many of the assumptions made. However, it is 
strongly believed that a further and closer look should help remove these deficien- 
cies and render the above approach in modeling an important addition to all other 
commonly adopted methods of analysis. 

Acknowledgment 

The authors thank the Purdue Research Foundation for supporting this work 
under grant number XRO340. 

Literature Cited 

1. Cohon, J. L. and D. H. Marks. "A Review and Evaluation of Multiobjective 
Programming Techniques," Water Resources Research, 11(2), 208-220, April 
1975. 

2. Eaton. D. J. and W. J. Steele (Editors), "Analysis of Grain Reserves, A Pro- 
ceedings", Report No. 634, Economic Research Service, U.S.D.A., August 
1976. 

3. Eaton. D. J., "A Systems Analysis of Grain Reserves," Technical Bulletin No 
1611, U.S.D.A., 1980. 

4. Houck. M. H. and D. J. Eaton. "Water and Grain Storage Systems: A 
Technology Transfer," International Conference on Operations Research in 
Agriculture and Water Resources, Jerusalem, November 1979. 

5. Poston, T. and I. Stewart, "Catastrophe Theory and its Applications," Pitman, 
London, 1978. 

6. Samuelson, P. A., "Foundations of Economic Analysis," Harvard University 
Press, Cambridge, 1961. 

7. Zeeman, E. C, "On the Unstable Behaviour of Stock Exchanges," Journal of 
Mathematical Economics, 1(1974), 39-49, 1974. 



Reservoir Operating Rules 

Mohammad Karamouz and Mark H. Houck 

School of Civil Engineering 

Purdue University, West Lafayette, Indiana 47907 

Introduction 

This study considers a deterministic optimization model to determine general 
reservoir operating rules. An algorithm that cycles through a deterministic 
dynamic program, a regression analysis and a hydrologic simulation has been 
developed. At each iteration the reservoir release becomes more constrained and 
the general operating rule becomes more refined. This work is the logical extension 
of Young's (3) study to refine the general operating rules by repeating the process 
with one extra constraint on the dynamic program; namely, release must be within 
some specified percentage of the release defined by the previously defined general 
operating rule. 

This algorithm was tested using annual streamflow data for the Gunpowder 
River, Maryland, the Osage River, Missouri, and the Blacksmith River, Utah. In all 
cases, significant improvements in operations of the reservoirs have resulted from 
use of the algorithm. In the remainder of this paper the algorithm is described, 
some results are presented, and finally a discussion of the results and a set of con- 
clusions are given. 

Algorithm to Refine Operating Rules 

A discrete dynamic program has been formulated to provide the optimal 
operating policies for a single reservoir over a finite time horizon of T years. The 
reservoir inflows for all T years and the reservoir capacity are assumed to be 
known. The objective function is to minimize the total losses caused by very high or 
very low streamflows over the T year time horizon. This can be expressed by 

T 
Minimize E Loss (R f ) (1) 

t=l 

The loss function (Loss (R t )) used in this investigation responds to reservoir 
outflow (R t ) only. As the deviation of outflow or release above or below some accep- 
table range increases, the losses associated with the release also increase. 

This problem is subject to some physical constraints. The continuity or mass 
balance at the reservoir should be preserved at all times. The discrete version of 
mass balance can be written as: 

St+l-St + Rt^t t=l,2,...,T (2) 

where: I t = inflow volume during year T 

S t = storage volume at the beginning of year t 

R t = outflow volume from the reservoir during year t 

Depending on the reservoir system there may also be restrictions on the maximum 
and minimum allowable outflows and storages. 



R max > R t > R min t = 1, 2, ..., T (3) 

t — l — t 



S max > S f > S min t = 1, 2 T (4) 

t — l — t 

The solution of the dynamic program comprises optimal outflows R t and optimal 

230 



Engineering 



231 



storages S t for the T year time horizon. These optimal storages and releases are 
dependent on the particular sequence of inflows used in the dynamic program and, 
therefore, do not define operating rules for the reservoir. 

However, the optimal outflows R t can be regressed against the optimal 
storages S t and inflows I t to provide general operating rules. 

R t = al t + bs t + c (5) 

where: a, b, and c are constants defined by a multiple regression. The consistency 
and the efficiency of these rules can be tested by simulation of the rules in opera- 
tion of the reservoir over a long time horizon. 

To refine an operating rule, the dynamic program can be used. If an operating 
rule already available would require the release at time t to equal ^ then the 
allowable release in the dynamic program can be bounded between plus and minus 
some percentage of this general operating rule: 

(6) 



(1 - BOUND) ** t < R t < (1 + BOUND) \ 



where: BOUND = fraction of general operating rule that actual outflow may devi- 
ate from the rule in the dynamic program. 

The solution of this dynamic program can be used to define a new, refined, general 
operating rule. This cycle — dynamic program, regression, simulation, dynamic pro- 
gram, ... — can be continued with the value of BOUND decreasing until it equals 
zero. 

Case Study 

In order to test the algorithm and its ability to produce good general 
operating rules, several cases were studied including different reservoir sizes and 
different streamflow sites. The streamflow sequence used in the dynamic program 
was the historical streamflow record. The simulation model was constructed for a 
1,000 year time horizon. The streamflow sequence used in the simulation model 
was generated by the autoregressive (AR) or auto-regressive-moving-average (AR- 
MA) model with the best AIC value (1). 

The loss function used for all cases was a discontinuous exponential function. 
As the outflow increases from zero to a safe range of outflow, this function ex- 
ponentially decreases to zero; and after a safe range, it increases exponentially. 

Table 1 is a sample result which clearly expresses the significant value of the 
algorithm in selecting reservoir operating rules. 

Table 1. Blacksmith River, Reservoir Capacity Equals 1.1* l(fim s 



Dynamic Program Results 



General Operating Rules 



Simulation Results 



BOUND 



losses/year 



% Time % Time 

Reservoir Reservoir 
losses/year Empty Filled 



0.36 


12.299. 


0.441 


0.066 


59.343. 


0.521 


74.932. 


19.0 


17.6 


0.18 


16.606. 


0.478 


0.053 


55,769. 


0.655 


76.019. 


18.0 


17.6 


0.09 


26,285. 


0.524 


0.015 


52.848. 


0.774 


78.901. 


19.0 


18.0 


0.06 


31.619. 


0.553 


0.106 


44,238. 


0.790 


75.035. 


12.9 


13.2 


0.03 


47.546. 


0.593 


0.262 


30261. 


0.914 


74,122. 


3.9 


6.1 


0.01 


107.362. 


0.764 


0.677 


- 14.429. 


0.838 


107,756. 


0.0 


0.0 



232 Indiana Academy of Science 



Discussion of Results 

1. As the value of BOUND decreases, R/ (goodness of fit) increases in general. 
This is expected because as BOUND goes to zero all releases are required to 
be closer and closer to the regression line. 

2. As the BOUND decreases, the losses/year in the dynamic program increase. 
Once again this is expected because as the value of BOUND decreases, there 
is a tightening of the constraints of the dynamic program and, therefore, the 
objective function suffers more. 

3. As the value of BOUND decreases to zero the general operating rule changes 
drastically. In general the value of c decreases and the values of a, and b in- 
crease. These changes signify a shift in the rule from a relatively constant 
release to one that depends more on inflow and storage. 

4. The loss/year from the dynamic program is less than the loss/year from the 
simulation. This is due to the optimal selection of outflows in the dynamic pro- 
gram. 

5. As the BOUND decreases from infinity to zero, in most of the cases, the 
simulation's loss/year first decreases then increases. 

6. In the simulation, the percentage of time that the reservoir is either filled or 
empty (implying that the actual outflow deviated from the operating rule) is 
inversely correlated to the losses/year. In all test cases the minimum percen- 
tage of filled or empty time occurred within minimum BOUND value of 0.01. 

Choosing the best operating rule for a reservoir involves many criteria, com- 
parisons and trade-offs. Two criteria that are explicitly considered here are the 
simulation's losses/year and the percentage of time that actual outflow is unequal 
to the outflow defined by the operating rule (which equals the percentage of time 
that the reservoir is filled or empty). The assumption is that each criterion should 
be minimized. This analysis demonstrates that the two criteria are conflicting and 
it provides the information needed to choose that rule with the best compromise 
between the two criteria. 

Rules with a form other than the linear one described here could also be used. 
However, as the form of the rule becomes more complex (e.g. non-linear), the com- 
putational effort of the algorithm increases. Bhaskar and Whitlach (2) have found a 
simple, linear operating rule is as good as or better than the more complex rules in 
many cases. 

Conclusion 

An algorithm to generate reservoir operating rules has been proposed and 
tested. The algorithm is easy to use and each component of the algorithm (deter- 
ministic dynamic program, multiple regression, simulation) is relatively simple and 
well documented in the literature. The algorithm can be used for complex multiple 
reservoir systems and for seasonal or annual operation of the reservoirs. All the 
test cases with different reservoir capacities and different streamflow 
characteristics show significant improvements in the operating rules are possible 
when the operating rules are refined using the proposed algorithm. 

Acknowledgment 

This material is based upon work supported by the National Science Founda- 
tion under Grant No. CME7916819. 



Engineering 233 

Literature Cited 

1. Akaike, H., A New Look at the Statistical Model Identification, Automatic 
Control, Vol. AC-19, No. 6, December, 1974, pp. 716-723. 

2. Bhaskar, N. R., and Whitlach, E. E., Jr., Derivation of Monthly Reservoir 
Release Policies, Water Resource Research, Vol. 16, No. 6, December, 1980, 
pp. 987-993. 

3. YOUNG. G. K., Jr., Finding Reservoir Operating Rules, Journal of the 
Hydraulics Division, ASCE, Vol. 93, No. HY6, November, 1967, pp. 297-321. 



ENTOMOLOGY 

Chairman: Michael Sinsko 
Indiana State Board of Health, 1330 West Michigan Street, Indianapolis, Indiana 
46206 

Chairman-Elect: David K. Reed 
Fruit and Vegetable Insect Research Laboratory, USDA-ARS Box 944, Vincennes, 
Indiana 47591 



ABSTRACTS 

Notes on the Biology and Oviposition of the Twobanded Japanese Weevil 

{Callirhopalus bifasciatus Roelofs). William E. Chaney, Donald L. Schuder, and 

David K. Reed, USDA-SEA, Vincennes, Indiana 47591. A study was conducted 

on the biology and oviposition habits of the twobanded Japanese weevil, 
Callirhopalus bifasciatus Roelofs. A number of plant species were examined to 
determine feeding preference of this species. The effects of temperature, humidity, 
and substrate on oviposition and egg hatch were examined. A number of general 
observations on the behavior and biology of both the adult and larvae were made. 

The number of pods produced by a given weevil was influenced by such en- 
vironmental conditions as amount of contact with the substrate, temperature, and 
humidity. The nature of the substrate itself, natural or artificial, significantly 
affected the number of eggs per pod. 

Oviposition, as measured by egg pod formation, was affected more greatly by 
humidity than temperature at the ranges examined. The number of eggs produced 
by an adult was not affected by the nature of substrate, since the number of pods 
produced was inversely proportional to the number of eggs per pod. Temperature 
and humidity did affect the number of eggs produced since the number of pods 
changed, but the number of eggs per pod did not. The percent of egg hatch was not 
significantly affected by either temperature or humidity, but the duration of the 
egg stage was affected. 

Laboratory and Field Evaluation of Oviposition Preference in Toxorhynchites 
rutilus rutilus. A. Yumi Clemons and Raymond J. Russo, Department of Biology, 
Indiana University-Purdue University at Indianapolis, Indianapolis, Indiana 

46205. The oviposition preference of Toxorhynchites rutilus rutilus (Theobald) 

was examined through the use of various oviposition substrates: distilled water, 
food water, larval water, and larval water with prey. Replicated 5 gal. cage experi- 
ments using 15 females showed a significant difference in the number of eggs laid 
on each type of substrate. Similar results were obtained when females were releas- 
ed in isolated trees in the field which were equipped with artificial treeholes con- 
taining the same type of water treatments. A distinct preference was shown for 
water which presently contains or once had living prey larvae. This enables Toxor- 
hynchites to select oviposition sites which have a high probability of containing 
food for its offspring. 

Clinical Variation of Temperature Dependent Survivorship in the Mosquito A edes 
sollicitans. Rosemarie Funkhouser, Department of Anatomy, Indiana University 
Medical Center and Raymond J. Russo, Department of Biology, Indiana 
University-Purdue University at Indianapolis, Indianapolis, Indiana 46205. 

234 



Entomology 235 

Survivorship of three strains of Aedes sollicitans from a north-south cline was 
determined at three constant temperatures of 22°, 25° and 28°C. Results were 
analyzed both graphically and statistically. The northern strain, Cedar Island, had 
better survivorship at temperatures of 22°C and 25°C. The intermediate latitude 
strain, Ft. Jackson, had better survivorship at 28°C and poorer at 22°C. The most 
southern strain, Ruskin, showed no significant difference in survivorship at the 
three separate temperatures. 

Spring-tailed Insects of the Genus Proisotoma, subgenus Appendisotoma, from 
Manlove Woods. John W. Hart, Division of Education, Indiana University East, 
Richmond, Indiana 47374. Collections of Collembola (1974 to date) from this uni- 
que woods (see A. A. Lindsey, et al. Natural Areas of Indiana and Their Preserva- 
tion) have contained several species of this genus/subgenus. Proisotoma (A.I 
vesiculata was the subject of a previous paper. One species currently under study 
appears to be Proisotoma (A.) bulbosa Folsom. If this is true, Folsom's description 
was based on ecomorphic specimens; and since the Manlove Woods material con- 
tains both normal and ecomorphic forms, it will be possible to fully describe this 
species. The ecomorphic form of a second species closely resembles D. L. Wray's 
published description of Frisonia veca. It would be desirable to study types of his 
species. An additional species is similar to our normal "veca" specimens, but it has 
only been taken in normal form. Additional study of the Proisotoma (A.) is in- 
dicated. 

Oviposition and Larval Development of Troxorhychites brevipalpis (Diptera: 
Culicidae) at Selected pH Levels. Sue Henderson and Robert R. Pinger, Depart- 
ment of Physiology and Health Science, Ball State University, Muncie, Indiana 

47306. A study was conducted to determine the effects of pH on oviposition, 

larval development, pupation and emergence success of Toxorphynchites 
brevipalpis. For the oviposition preference study ten male and ten female T. 
brevipalpis mosquitoes were placed in each of 8 1' x 1' x 1' cages. Four oviposition 
sites containing rainwater adjusted to pHs of 4, 5, 6 and 7 were placed in each cage 
and rotated daily to nullify the effect of lighting and air movements. Eggs were 
counted daily and tabulated by pH and position of site. This experiment was 
replicated four times, giving a total of 32 replications. No oviposition preference 
was indicative with regard to pH but differences were noted in the preference ex- 
hibited for site location within cages. 

In the second phase of the study, 30 first instar larvae were individually raised 
in glass containers at each of 5 pH levels (3, 4, 5, 6, 7). Larvae were fed ad libitum 
with larvae of Aedes triseriatus . Larvae were examined each day and mortality, 
dates of molting, pupation and emergence were recorded. Data were examined for 
differences in rates of development and success of molting, pupation and 
emergence. 

Mortality was greatest at pH 3. Development appeared to be most successful 
at pHs 4 and 5. Differences in development rates in different instars were noted. 

Effectiveness of the Ovitrap for Monitoring Culex Populations. Bridget Hoban. 
and George B. Craig. Jr.. St. Joseph County Mosquito Abatement Program, 
Department of Biology, University of Notre Dame, Notre Dame, Indiana 

46556. In an attempt to monitor populations of Culex pipiens and Culex res- 

tuans, Culex ovitraps were used throughout the county. These ovitraps were 
designed to collect egg rafts from ovipositing females. The trap was a 5-liter plastic 
bucket with a cover in place but ajar to leave a 1 inch opening; the trap contained 



236 Indiana Academy of Science 

water and cow manure in a cheesecloth sack. On May 21, 25 ovitraps were 
distributed throughout the county, with 11 of them placed close to a light trap. Egg 
rafts were collected on Monday, Wednesday and Friday of each week until 23 June. 
On 27 June the ovitraps were replaced with fresh ovitraps. Egg rafts were col- 
lected and identified to species as before. However, beginning July 9, only 20 rafts 
per site were identified because some traps were producing up to 100 egg rafts and 
logistics for rearing became impossible. Altosid was added to all the traps. The 
ovitraps were replaced again in 14 July. At the five most productive sites, a second 
ovitrap lacking Altosid was added. This was to determine whether Altosid had an 
influence on ovipositing females. These ovitraps were discontinued on 6 August. 
Over the summer, 8,353 egg rafts were collected. Culex restuans was predominant. 
There were no Culex pipiens collected until 27 June. At the end of the experiment 
on August 6, the proportion of Culex pipiens had increased to only 35%. The 
Altosid was effective in preventing adult emergence; and from results of the 
Altosid comparison tests, the Altosid did not significantly reduce the effectiveness 
of the traps. The total numbers of rafts collected per trap collected corresponded 
with the total number of mosquitoes caught out of the light traps. The use of the 
manure attractant oviposition traps has been a very successful method of monitor- 
ing Culex populations in St. Joseph County. By standardizing the traps, and by ad- 
ding Altosid to prevent adult emergence, we have developed a good tool. 

The Burrowing Mayflies (Ephemeroidea) of Indiana. John Keltner and W. P. Mc 
Cafferty, Department of Entomology, Purdue University, West Lafayette, In- 
diana 47907. Twelve of the thirty species of burrowing mayflies have been 

recorded for Indiana, representing four families and six of the nine U.S. genera. 
General information was sought regarding geographic and phenological distribu- 
tion and aquatic micro-habitat preferences for all Indiana species along with detailed 
burrowing and feeding behavior of larval exemplars for all genera. Larval 
Potamanthus inhabits lotic environments, clinging to the undersides of rocks and 
other surface substrates, however, larvae of all other genera live within bottom 
substrates at various depths and construct various subterranean burrows. 
Ephemera and Ephoron, in Indiana, are found in a wide range of lotic and more 
rarely lake substrates, from cobble-sand-gravel to finer sediments. Hexagenia, in 
Indiana, occurs in soft, often organically enriched, silt bottoms in both lentic and 
lotic environments, whereas Tortopus and Pentagenia are adapted for burrowing 
in compacted clay bottoms and/or submerged banks of large rivers. Most genera 
are widely distributed in the state. Tortopus and Pentagenia, however, have been 
taken only on the lower Wabash River. Pentagenia was apparently previously 
more widespread in the larger rivers. Ephoron and Tortopus are univoltine with 
extremely synchronized and short emergence periods. Hexagenia, Pentagenia, and 
Ephemera have more complex life histories with overlapping generations and 
more prolonged population hatches. Potamanthus is univoltine or possibly requires 
two years for development with a very prolonged summer emergence period. 

Effects of pH on Oviposition Preference and Larval Development of Mosquito 
Species Aedes triseriatus. Stephen R. Madigosky, Robert R. Pinger and Horst F. 

Siewert, Ball State University, Muncie, Indiana 47306. A laboratory study was 

conducted to determine preference for pH levels of water by gravid female mos- 
quitoes for oviposition and to evaluate effects of low pH levels on larval develop- 
ment of Aedes triseriatus. Oviposition preference was assessed by allowing F 3 
generation females to lay eggs on one of six available oviposition sites. Seventy 
Aedes triseriatus females were placed in separate pint containers. Each container 



Entomology 237 

accommodated six glass vials with rainwater, which was adjusted to one of six dif- 
ferent pH levels (2 through 7). All vials contained a thin strip of balsa-wood which 
provided a medium for oviposition. The balsa-wood strips were soaked at pH levels 
2 through 7, 21 days prior to use. Female mosquitoes were retained in the con- 
tainers for 10 days. Very little oviposition preference was established between pH 
4 through 7, however, oviposition sites at pH levels 2 and 3 were avoided. 

Larval development was assessed by allowing 50 first instar larvae to develop 
in separate glass containers at 5 different pH levels 3, 4, 5, 6, 7. and a control. The 
time from hatching to pupation, and from pupation to emergence of adults were 
monitored. Mortality was greatest at pH levels 3 and 4. 

Effect of Prey Density on the Use of Toxorhynchites rutilus as a Biological Control 
Agent. Raymond Russo and Kimberly Raubenheimer, Department of Biology, In- 
diana University-Purdue University at Indianapolis, Indianapolis, Indiana 

46205. Toxorhynchites rutilus rutilus (Theobald) larvae were reared on Aedes 

aegypti (Linnaeus) prey larval densities of 5, 10, 25, and 50. Rate of development 
and amount of prey consumed were measured. 

Tx. r. rutilus larvae offered 5, 10, 25, and 50 prey larvae daily, required 23.4, 
21.5, 20.6, and 19 days, respectively, to complete development through emergence. 
As prey density increased, the number of prey consumed increased. Tx. r. rutilus 
larvae offered 5, 10, 25, and 50 prey daily consumed an average of 73, 106, 163, and 
231 prey, respectively, during their larval development. A prey/predator conver- 
sion efficiency was determined by obtaining dry weights of prey and estimating the 
dry weight of predatory larvae. 

The results of this experiment show that the rate of development of Tx. r. 
rutilus can be controlled by the quantity of prey available. 

Home Garden Insecticide Use in Indiana. Alan C. York and Ronald D. Gardner 
Department of Entomology, Purdue University, West Lafayette, Indiana 

47907. A survey was conducted of 2800 home gardeners in Indiana. This survey 

consisted of face to face interviews of equal numbers of people in rural, small town 
(less than 5,000), medium town (5,000 to 50,000), and large city (greater than 50,000) 
areas. It was found that 54% of households surveyed used insecticide on 
vegetables or fruit. Information was obtained on 57 brands of 21 different active in- 
gredients. Carbaryl was used most frequently, 44% of the uses. More than 50% of 
the uses on cucumber, cantaloupe, squash, pepper, beans, sweet corn, and potato 
were of this compound. Rotenone was second most frequently used, 29%. Methoxy- 
chlor (6.85%), malathion (6.69%), diazinon (5.6%), and pyrethrum (2.1%) comprised 
the majority of the remainder. Of the remaining compounds, only chlordane was 
used more than 1.0% of the time. Dusts were used most frequently (76.1%) follow- 
ed by liquid, wettable powder, aerosol, and granular formulations. Most people 
(53.8%) said that they purchased insecticides by common name rather than by 
brand name; however, Ortho products were purchased 61 + % of the time. Use of 
insecticide was slightly, but positively correlated with age, years of experience, 
and the size of garden. 



Treehole Productivity for A edes triseriatus in Northern Indiana, 1975 vs. 1979 

Stephen L. Durso and George B. Craig, Jr. 

Vector Biology Laboratory 

University of Notre Dame, Notre Dame, Indiana 46556 

Introduction 

The treehole mosquito, Aedes triseriatus (Say), has been known for about 
eight years as the major vector of LaCrosse Encephalitis in the Midwest (6). 
LaCrosse, a member of the California group of encephalitis viruses, is responsible 
for serious illness in hundreds of children in the Midwest; as recent studies show, it 
may be far more prevalent in human populations than previously thought (3). 
Therefore, the location and control of this mosquito is important. 

Aedes triseriatus breeds primarily in treeholes and rot cavities. However, it 
has been found increasingly more often in a wide variety of man-made artificial 
habitats, such as tires, rain gutters and birdbaths. Discarded tires rank as the most 
common urban source. Several factors make detection and surveillance a difficult 
endeavor. Conventional surveillance methods for mosquitoes fail to detect signifi- 
cant numbers because Aedes triseriatus is not attracted to light or carbon dioxide; 
moreover, it will not enter live-baited traps (7). The inaccessibility of treeholes has 
made larval surveys difficult. To date, the best method for detecting the presence 
of Aedes triseriatus has been through the use of a modification of the Fay oviposi- 
tion trap, in which a blackened aluminum can containing water and a balsawood 
strip is placed in woodlots to monitor female egg deposition (1). 

At present, effective control is impossible. Several control attempts, including 
the closing of all basal treeholes (2), have resulted in inconsistent results. The inac- 
cessibility of treeholes has led many to suggest biological control measures to 
reduce population levels. In order to evaluate the effectiveness of any control 
measures, a more efficient method of surveillance must be found. 

Kramer's Woods is an isolated woodlot of roughly 10.1 hectares in size in St. 
Joseph County, Indiana. The woodlot consists primarily of secondary growth Black 
and White Oak, creating an abundance of basal treeholes. The surrounding corn 
fields and pine forests serve to make Kramer's Woods, in effect, an ecologically 
isolated island in which to study the endemic population of Aedes triseriatus. 

For these reasons, Dr. Michael Sinsko chose Kramer's Woods to conduct 
Aedes triseriatus population studies in 1975 (5). His survey consisted of locating all 
the treeholes within the woods and tabulating the size, location, and tree type of 
each. To monitor the population of Aedes triseriatus, he made weekly pupal counts 
of each treehole from mid-March to mid-September. His method involved emptying 
the contents of each treehole using a Foley turkey baster, sexing and isolating the 
pupae into emergence containers, and replacing the contents back into the 
treehole. He determined both the seasonal production of A edes triseriatus for each 
treehole and the overall production of the woodlot. 

To determine whether or not there is fluctuation in productivity from year to 
year, or whether the physical makeup of the woods (i.e., number and location of 
treeholes) changes yearly, a similar study was undertaken in the summer of 1979. 

Materials and Methods 

The study site, Kramer's Woods, is located 9.6 km northwest of the University 

238 



Entomology 239 

of Notre Dame campus. The areas surrounding the woodlot remain relatively un- 
changed since 1975 with a conifer forest to the north, a cultivated field to the south, 
and open fields to the east and west, thereby maintaining the isolated character of 
the woods. 

All treeholes were located in early spring following periods of heavy rainfall, 
using a series of north-south and east-west transect searches. Criteria held for a 
positive treehole were the ability to hold water and the presence of biological life. 
Each treehole located was flagged, marked with an aluminum tag, and its size, loca- 
tion, and tree type were recorded (Figure 1). 




# 306 



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*203 


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206 






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Figure 1. Map of Kramer's Woods, showing treehole distribution in 1979. 



240 Indiana Academy of Science 

In order to study the Aedes triseriatus population of the woodlot, an 
emergence trap was employed. The emergence traps, modified from Yates (8), were 
constructed by stapling tubular stockinette (American Hospital Supply) around the 
hole. A collection vial equipped with an inverted paper funnel at its opening was at- 
tached to the distal end of the stockinette with a rubber band. The distal end was 
secured to a nail or stick so that the collection vial was perpendicular to the 
treehole. 

Emergence traps were set on all treeholes between 900 and 1100 hours, and 
were collected during the same period the following day. Adults were collected two 
days per week from June 26 through July 18, three days per week from July 24 
through August 31, and two days per week from September 5 through September 
20. Adults located in the traps each day were sexed, counted, and released back 
into the woodlot. It was hoped that this trapping system would yield a represen- 
tative sample from which total emergence could be estimated. 

All rainfall data were obtained from the National Weather Service Office of 
South Bend, Indiana, Located approximately 2.5 km from Kramer's Woods. 

Results 

The map of the location of the treeholes found in Kramer's Woods in 1979 
shows, as it did in 1975, that the treeholes appear to be clumped throughout the 
woods with the greatest number being found in the east and southeastern direc- 
tions (Figure 1). 

The 1979 survey yielded 104 treeholes, as compared to 108 found in 1975 
(Table 1). On closer inspection, only 53 treeholes found holding water in 1979 were 
also recorded in the 1975 survey. This represents a mean yearly treehole turnover 
of 12.2 trees. The percentage of treeholes in the two types of oak was very close 
with roughly twice as many Black Oak showing treeholes as White Oak in each of 
the two years. The examination of the physical parameters of all the treeholes 
seemed to show a difference between the two years. However, when examined for 
only those treeholes known to be common to the two surveys, there is very little 
change in the physical measurements. 

The total production of Aedes triseriatus was relatively close for each of the 

Table 1. Analysis of treeholes in Kramer's Woods, 
St. Joseph Co., Ind., 1975 vs. 1979 

Treehole Year of Analysis 

character 1975 1979 

Total number 108 104 

Species composition in % 
Black Oak 
White Oak 

Dimensions of hole in cm 
All treeholes 
Length 
Width 
Area (cm 2 ) 

Common treeholes (N = 53) 
Length 
Width 
Area (cm 2 ) 

Annual rate of treehole turnover = 12.2 



67 


63 


33 


37 


13.0 


11.3 


8.1 


6.9 


116.1 


87.3 


12.7 


12.1 


7.5 


7.0 


102.2 


97.3 



Entomology 241 

Table 2. Treehole productivity for A edes triseriatus in 
Kramer's Woods, 1975 vs. 1979 





Adults 
produced 


Year of analysis 




1975 1979 


Female 

Male 

TotaJ 




1790 2207 
2438 1393 
4228 36QQ. 


Sex Ratio, 9:0" 




1:1.4 1:0.63 



two years, differing by only 600 individuals (Table 2). The difference in the total 
numbers may be attributed to the relatively low number of males in 1979. There 
was roughly one-half the number of males in 1979 as there was in 1975, while a 
slight increase in females was seen in 1979. This is responsible for the reversal of 
the overall sex ratio from favoring males in 1975 to favoring females in 1979. 

Upon examination of the seasonal emergence of both males and females for 
the two years, there appears to be a striking dissimilarity between the two years 
(Figure 2). However, the general pattern of emergence is the same for each year. In 
each of the years there is a favoring of males over females in early summer, 
gradually diminishing to a 1:1 ratio in mid-summer. This is followed by a late sum- 
mer favoring of females over males. Conspicuously different in the seasonal 
emergence patterns is the degree to which each of the years favors production of 
one or another of the sexes in either early or late summer. The high sex ratio favor- 
ing males in mid-June of 1975 is not duplicated to such a great extent in 1979. Con- 
versely, there is a greater favoring of females in early September during 1979 than 
was seen during 1975. During these few weeks in 1979, there were three times 
more females than males produced within the same period. 

The apparent shift in the 1979 emergence is not an environmental event, but 
rather a reflection of the monitoring method. The 1975 survey utilized pupal 
counts, while the 1979 survey employed the subsequent adult emergence monitor- 
ing techniques, thereby explaining the shifting of the 1979 emergence pattern in 
relation to 1975. 

In an attempt to explain this great difference in the number of Aedes 
triseriatus while the overall seasonal pattern was consistent, the seasonal produc- 
tion was examined in relation to the weekly rainfall. The correspondence between 
the rainfall and mosquito production shows up very clearly in the 1975 survey 
(Figure 3). The build-up of rain in late May and early June accounts for the peak of 
pupae produced in mid-June. This is followed by a fluctuating rainfall period that 
corresponds to moderate pupal production; a period of great rainfall in early 
August accounts for the late August peaks. 

The 1979 data show a moderate amount of rainfall in May, with a build-up in 
early June accounting for the production of adults in early July (Figure 4). This is 
followed by a relatively dry period with a reduction of emergence. The rapid build- 
up of rainfall in late July culminates in over 4.2 inches of rainfall during one week in 
early August. It is during this time that a tremendously high number of treeholes 
is being filled, with some treeholes beginning to produce larvae for the first that 
season. This high amount of rain in early August is responsible for the enormous 
number of adults produced in late August and early September, at a time when a 
good deal of the treeholes were already dry for the season in 1975. 

A ranking of the contribution by each individual treehole to the productivity 



242 



Indiana Academy of Science 



600 



200 



(1975) 



♦Female 
•Male 




May June July August September 

FIGURE 2. Emergence pattern for each sex of Aedes triseriatus 
' for the years 1975 and 1979. 



of the woodlot (Figure 5) reveals a very similar relationship in each year. Of 
notability is that approximately one-third of the treeholes in each year never pro- 



Entomology 



243 



duce any Aedes triseriatus (37% in 1975 and 35% in 1979). 

Sinsko reported that in 1975 over 83% of all pupae were produced by only 
22% of the treeholes, with at least two treeholes producing over 500 individuals 
apiece. A similar event was seen in 1979. However, in 1979, 83% of all adults were 
produced by 31% of the treeholes, and not more than one treehole approaches even 
one-half the production of the most productive treeholes in 1975. This illustrates 
that production of Aedes triseriatus was more evenly distributed throughout the 
treeholes in 1979 than in 1975. 

Discussion 

The clumped distribution of treeholes in Kramer's Woods may simply be a 
reflection of the overall distribution of all trees within the woodlot. Sinsko (5), using 



(1975) 



♦Female 
•Male 




5 



_ 3 0- 

C 
CO , 

QC ' 
>> 

0) 
CD 



n 



a 



a 



n 



fl - nn 



May June July August September 

Figure 3. Aedes triseriatus emergence and weekly rainfall for 1975. 



244 



Indiana Academy of Science 



Female 
Male 




r " " ■"■ ■" i » 

May June July August September 

Figure 4. Aedes triseriatus emergence and weekly rainfall for 1979. 



a point-quarter analysis, was able to divide the woods into two distinct area based 
upon the abundance of saplings and mature trees. His two areas (treehole area and 
scrub area) correspond roughly to the clumping of the treeholes and the treehole 
sparse areas. Sinsko calculated that there were 58% Black Oak and 40% White 
Oak occurring in the treehole area. Therefore, the percentage of each type of oak 
with treeholes found in either year would not reflect a tendency for one type of oak 
to form treeholes over the other. The comparison of physical measurements of the 
treeholes shows that there are distinct populations of treeholes in each of the two 
years, while the actual treeholes themselves changed very little in size during this 
period. 



Entomology 



245 




50 100 

Ranked Treeholes 

Figure 5. Ranking of treehole productivity for Aedes triseriatus in Kramer's 
Woods for the years 1975 and 1979. 



246 Indiana Academy of Science 

The fact that a relatively small number of treeholes endured through both 
studies, while the total number of treeholes was constant, further demonstrates 
the fluctuating relationship of the treeholes within the woodlot and the stability of 
the entire woodlot. This is an excellent example of the difficulties involved in the 
control of Aedes triseriatus based upon the knowledge of the location of all breed- 
ing treeholes within a woodlot. As treeholes are added or become no longer capable 
of holding water each year, control programs based upon location of treeholes 
would become increasingly difficult without costly searches for new treeholes be- 
ing done each year. 

The seasonal emergence pattern of an early favoring of males gradually 
switching over to a late season favoring of females in Kramer's Woods has been 
documented for several years (4,5). Shroyer (4) has suggested several possible 
mechanisms for the seasonal favoring of one sex or the other based upon a number 
of environmental conditions and genetic effects. One of his proposed mechanisms 
states that males seem to exhibit an earlier cessation of overwintering diapause 
than females, and therefore are seen in much greater numbers early in the sum- 
mer. Males also tend to have a greater percentage of egg hatching at longer 
photoperiods (early and mid-summer conditions), with the percentage hatch 
gradually diminishing as photoperiod shortens (late summer conditions). On the 
other hand, females exhibit a greater percentage of egg hatching at shorter 
photoperiods, thereby explaining their overwhelming numbers later in the sum- 
mer. Shroyer has further stated that, at a certain critical photoperiod, both sexes 
undergo a dramatic increase in the percentage of eggs that enter into an over- 
wintering diapause, and, therefore, no more hatching of eggs takes place. For 
Kramer's Woods, Shroyer has calculated the critical photoperiod for females to be 
around August 10th, with that of males occurring as much as a week earlier. 

The examination of emergence patterns for the years 1975 and 1979 support 
Shroyer's proposed mechanism, as far as the actual production of adults is concerned. 
In both years, the switchover of emergence from males to females has been ex- 
hibited. Of all the years that emergence in Kramer's Woods has been studied, 1979 
is the first year that the overall seasonal sex ratio has favored females. This is at- 
tributed to the enormous peak of females in early September. This peak of females 
could be explained, using Shroyer's proposed mechanism, as being due to the great 
amount of rainfall in early August. If this rainfall occurred before the initiation of 
the critical photoperiod for females and just after the initiation of the male 
diapause, it would have had the effect of filling a greater proportion of the 
treeholes with water, and the hatching out of a number of eggs, the majority of 
which were females. In August of 1975, there was a good amount of rainfall, but 
there is not as great a peak of females produced from it. This may be due to the fact 
that the rains in that year may have been just after the initiation of the critical 
photoperiod for both sexes, and therefore a good deal of the eggs of each sex had 
already entered diapause. 

Remaining to be explained is the lack of a strong favoring of males in early 
summer of 1979. It may be that the winter environmental conditions, or the spring 
temperatures or rains, were not optimal for the hatching of eggs. 

The heavy rains in early August of 1979 had the effect of filling a number of 
previously unproductive treeholes, thereby distributing the production of Aedes 
triseriatus throughout a greater number of treeholes. This is seen in Figure 5, in 
which a greater number of treeholes were needed to produce the same percentage 
of total mosquitoes as a smaller number of treeholes accomplished in 1975. Of im- 



Entomology 247 

portant note here is the observation that a percentage of treeholes in each year 
never produced Aedes triseriatus at all, while a relatively small number of 
treeholes produced the majority of mosquitoes. This illustrates the importance of 
better knowledge of the physical, chemical, and environmental parameters of 
treeholes that attract ovipositing females or that enable larval development, in 
order to be able to target control measures based upon knowledge of treehole loca- 
tion. In the present study, no correlation was seen between either treehole size or 
volume and the production of Aedes triseriatus. 

Acknowledgement 

Funding was provided in part by NIH Research Grant No. AI-02753 and by 
The St. Joseph County Health Department. 

Literature Cited 

1. Berry, W. J., D. Fish, G. B. Craig Jr. 1979. The use of an ovitrap grid for 
measuring adult movement and population density of the treehole mosquito 
Aedes triseriatus. Proc. Ind. Acad. Sci. 89:208-209. 

2. Garry, C. E. and G. R. DeFoliart. 1975. The effect of basal treehole closure 
on suppression of Aedes triseriatus (Diptera:Culicidae). Mosq. News 
35:289-297. 

3. GRIMSTAD, P. R. 1980. An arbovirus serosurvey of 10,000 Hoosiers. Ohio Mosq. 
Cont. Assn. Proc. 9:7-8. 

4. SHROYER, D. A. 1979. Seasonal aspects of egg hatching in Aedes triseriatus 
(Say): Sex ratio distortion and diapause. Ph.D. Dissertation. University of 
Notre Dame, Indiana. 181 pp. 

5. Sinsko, M. J. and G. B. Craig, Jr. 1979. Dynamics of an isolated population of 
Aedes triseriatus (Diptera:Culicidae) I. Population size. J. Med. Entomol. 
15(2):89-98. 

6. Thompson, W. H., R. 0. Anslow, R. P. Hanson & G. R. DeFoliart, 1972. 
LaCrosse virus isolations from mosquitoes in Wisconsin, 1964-68. Am. J. 
Epidemoil. 38:245-253. 

7. Wright, R. E. and G. R. DeFoliart. 1970. Associations of Wisconsin mos- 
quitoes and woodland vertebrate hosts. Ann. Entomol. Soc. Amer. 
63(3):777-786. 

8. Yates, M. 1973. Emergence trap for sampling adult tree-hole mosquitoes. En- 
tomol. Mon. Mag. 109:99-101. 



Distribution of Aedes triseriatus (Say) in an Urban Area: Comparison of 
Two Survey Methods 

Lorraine Leiser 

Vector Biology Laboratory, Department of Biology 

University of Notre Dame, Notre Dame, Indiana. 46556 

Introduction 

Aedes triseriatus (Say) has been incriminated as the primary vector of La 
Crosse Encephalitis in the midwest (4). This mosquito is commonly found breeding 
in treeholes in forested areas. However, it also occurs in urban treeholes and other 
containers that are associated with man. There is increasing evidence that trans- 
mission of La Crosse Encephalitis occurs in urban and suburban areas as well as in 
more sylvan sites. 

We know very little about distribution and abundance of this mosquito 
because methods for monitoring populations are inadequate. Because their activity 
is diurnal, they are not attracted to light; therefore, light traps are not a reliable in- 
dicator of their presence or absence. Moreover, these mosquitoes will not readily 
enter holes or slots in animal-baited or CO2 traps. Therefore, traditional mosquito 
trapping methods are useless. 

Fay and Eliason (2) developed the ovitrap for use with Aedes aegypti. Their 
ovitrap consisted of a blackened 1 pint jar half filled with water; a wooden paddle 
partially submerged provided a surface for egg deposition. Loor and De Foliart (3) 
modified the Fay trap for use with Aedes triseriatus . They used a metal beer can 
painted with black enamel and lined with black cotton cloth. 

During the summer of 1980 we compared effectiveness of larval survey vs. 
ovitrap survey. Funding for this project was provided by the St. Joseph County 
Mosquito Abatement Project, St. Joseph Co. IN. Support was also provided by 
NIH Grant No. AI-02753. 

Methods 

The city of South Bend was divided into 48 sections. Using a system designed 
by Mr. John Stamper of the Office of Historic Preservation in South Bend, sections 
were determined on the basis of architectural type and period and formed 
neighborhood units. 

For the larval survey each section was surveyed by a team of workers visiting 
every 5th property along both sides of randomly selected streets. The survey of 
the entire urban area required more than three months to conduct, extending from 
May 5 to Aug. 15. Care was taken to cover the entire area of a section. Properties 
were classified as to structural type and use. These classifications were: Industrial- 
Commercial, Single Dwelling, Multiple Dwelling, Public Building, Public Lands 
such as parks, Woodlots, and Other. Breeding sites were classified as positive or 
potential. Positive sites were defined as those in which larvae were found. Poten- 
tial sites were those which, if wet, were judged capable of breeding mosquitoes. 
Positive sites were classified as follows: Artificial Container, Tires, Treeholes, 
Rubbish, Ornamental Lawn Containers such as birdbaths, Woodland Pools, Dit- 
ches, Lakes and Ponds, Streams and Creeks, Septic Overflow, Catch Basin and 
Other. Roof gutters were not included because of the difficulty in reaching these 
sites. Mosquitoes were classified into 4 groups: Anopheles sp., Culex sp., Aedes 
triseriatus and other Aedes. 

248 



Entomology 249 

A second survey was conducted using ovitraps to determine oviposition. A 
total of 94 ovitraps were in place throughout the city from June 18 to Sept. 27. The 
number of ovitraps varied from 1 to 5 with the size of the section. Modified Fay 
ovitraps (1) were placed in shaded concealed sites, often nailed to a tree trunk or 
fence post. Traps were examined five times during the three month period. The 
cans found to be missing or vandalized were either replaced or moved a short 
distance to a new location. Paddles with eggs were replaced with fresh paddles. 
Eggs/paddle were counted and eggs/paddle/day was calculated for each sampling 
period. 

Aedes hendersoni Cockerell is a sibling species, closely related to Aedes 
triseriatus and often occurring in the same habitat. No attempt was made to 
separate the two species in this work. 

Results 

During the larval survey, a total of 2386 properties were visited; of these 24 or 
1% were found to contain Aedes triseriatus. Of these properties found positive for 
the mosquito, a total of 29 sites were located, with Treeholes (23) the most 
numerous sites followed by Tires (5) and Ornamental Containers (1). Single Dwell- 
ing properties contained the highest proportion of treeholes (11) with Public Lands 
(5), Woodlots (4), Multiple Dwellings (2) and Industrial-Commercial properties (1) 
the remaining sites. Three tires breeding Aedes triseriatus were found on 
Industrial-Commercial properties while two more were located at Single Dwell- 
ings. The only Ornamental Container, found with Aedes triseriatus, a large foun- 
tain, also was located at a Single Dwelling. 

Aedes triseriatus breeding sites were found by the larval method in 16 or 
33% of the sections of South Bend (Figure 1). These sections were located in the 
North East and the South of the city. 

In the ovitrap survey, 94 ovitraps were placed throughout the city. A total of 
126 paddles from 55 different ovitraps were found to contain eggs during the 
course of the survey. Numbers of positive ovitraps for each sampling period were 
as follows: 

20 - July 3-10 

19 - July 22-24 

14 - Aug. 6-8 

26 - Sept. 2-5 

47 - Sept. 19-27 

Two periods of heavy egg production were evident. One occurred in July and 
the second in September. Eggs counted per sampling period were as follows: 



3414 


- July 3-10 


3141 


- July 22-24 


1469 


- Aug. 6-8 


3295 


- Sept. 2-5 


6750 


- Sept. 19-27 



18069 Total 

Eggs/paddle/day ranged from a high of 14 in July to a low of 2 in early September 
(Figure 3). 

Positive ovitraps were found in 32 or 66% of the sections. Aedes triseriatus 
was present throughout most of South Bend with the exception of the South West 
(Figure 1). 



250 



Indiana Academy of Science 




jjOvitrapSurvey 



: : ; : ." : :] Urban Survey 
Both Methods 



Figure 1. A comparison of sections found positive by Ovitrap and Larval 
(or Urban) survey in South Bend. 



Three main areas with heavy egg production were located by ovitraps. These 
areas are in the North, the East, and the South, South East of the city. One ovitrap 
contained eggs at all 5 sampling periods was found in each of these 3 areas as well 
as many traps found positive 3 or 4 times (Figure 2). 

Discussion 

The ovitrap method provided much more useful information about the 
presence of Aedes triseriatus in South Bend than did the larval survey. As Figure 
1 shows, the mosquito was found in most sections of the city. High numbers of 
positive ovitraps occurred in residential areas where many trees are present to 



Entomology 



251 



provide breeding sites (FIGURE 2). The South West sections of the city where few 
mosquitoes were found are heavily industrialized and trees and vegetation tend to 
be more sparce. 

From the ovitrap survey, it was also possible to gain information on popula 
tion trends during the season. The number of eggs/paddle/day can be correlated 
with the amount of rainfall in previous weeks (Figure 3). After significant rainfall 
in early August, egg production increased. It should be noted that no eggs hatch 
after August 10. Diapause in the egg stage is induced by the shortened day length, 
i.e., reduced light phase of the photoperiod. The number of ovitraps with eggs also 
rose during September, again indicating a rise in population from the previous 
weeks. 




Ovitrap Positive 

• 1 Time 

• 2 Times 

• 3 Times 
t 4 Times 

5 Times 
Ovitrap Negative 



Figure 2. Locations of ovitraps positive for Aedes triseriatus in South Bend. 



252 



Indiana Academy of Science 



50. 



40. 



> 30 
O 



| 20 

3 



10. 



Eggs/Paddle/Day 



Number of Ovitraps 
with Eggs ^ 



Number of New Ovitraps -W 
with Eggs 0* 




15 



10 



D 
5 w 



June 



July August September 



6-1 



5. 



3. 



-n-nnnflnn 



1 



n 



June 



July 



August September 



Figure 3. A comparison ofOvitrap Survey results for Aedes triseriatus with 
rainfall in South Bend, 1980. 



An advantage of the ovitrap method is that it used far fewer resources to gain 
a better picture of Aedes triseriatus distribution in South Bend. It took approx- 
imately 1/7 the man hours to perform that was needed by the larval survey 
(Ovitraps = 74hr., Larval Survey = 510hr.). It also resulted in far fewer miles being 
driven and thus a significant gasoline savings. Gasoline costs were about 1/4 those 
of the larval survey (Ovitraps = $55.00, Larval Survey = $217.00). In a time of in- 
creasing inflation this is an important point. 

The limitations of the larval survey method are many. Mosquito breeding 
sites are often concealed or inaccessible. The nature of this kind of survey makes it 
possible to visit a property, at most, only once or twice. As weather conditions and 
amount of rainfall vary, a site unproductive one week may be breeding mosquitoes 



Entomology 253 

the next. Also time and manpower limitations make it possible to survey only a 
small proportion of all the properties in each section, therefore missing sites. The 
larval survey does have the advantage that it pinpoints breeding sites. However, 
after ovitraps have indicated the presence of Aedes triseriatus in an area, a search 
can be made for these sites. This greatly narrows the areas to be searched. 

The ovitrap method identified many more sections positive than did the larval 
survey. It was able to monitor egg laying throughout the test period and pinpoint 
areas where heavy egg production was occurring. It also gave an estimate of 
population trends that the larval survey was unable to provide. Finally, it was far 
more efficient in the use of financial resources. 

Literature Cited 

1. Berry, W. J., D. Fish, and G. B. Craig Jr. 1980. The use of an ovitrap grid for 
measuring adult movement and population density of the treehole mosquito 
Aedes triseriatus. Proc. Indiana Acad. Sci. 80: 208. 

2. Fay, R. W. and D. A. Eliason. 1966. A preferred oviposition site as a 
surveillance method for Aedes aegypti. Mosq. News. 26:531-535. 

3. Loor, K. A. and G. R. DeFoliart. 1969. An oviposition trap for detecting the 
presence of Aedes triseriatus (Say). Mosq. News. 29:487-488. 

4. Thompson, W. H., R. 0. Anslow, R. P. Hanson, and G. R. DeFoliart. 1972. La 
Crosse virus isolations from mosquitoes in Wisconsin, 1964-68. Amer. J. Trop. 
Med. Hyg. 21:90-96. 



Indiana Pine Wilt Nematode Survey 

Philip T. Marshall 
Indiana Department of Natural Resources, Indianapolis, Indiana 46204 

and 

John J. Favinger 
Indiana Department of Natural Resources, Indianapolis, Indiana 46204 

Introduction 

The pine wilt nematode (or pinewood nematode, same common name) (Bur- 
saphelenchus lignicolus Maymiya and Kiyohara [Nematode: Tylenchida: Apelen- 
choididae]) is a serious disease to black pine iPinus thunbergiana Franco) and red 
pine (P. densiflora Sieb. and Zucc.) in Japan. As these pines are economically and 
aesthecially important in Japan, the scattered wilting and mortality to the forests 
and landscapes started research that proved B. lignicolus to be the causal agent (2). 
The research also found a long-horn beetle (Monochamus alternatus) to be the vec- 
tor (3). 

Dying pines, for which no cause of mortality could be found, have occurred in 
the United States. In Missouri in 1979, a sample from a wilting Austrian pine (P. 
nigra Arnold) was submitted to the Department of Plant Pathology at the University 
of Missouri. On the suggestion of a visiting Japanese professor of plant pathology, 
wood samples were soaked in water. Nematodes emerged from the wood and were 
identified as the pine wilt nematode (B. lignicolus). This was the first report of the 
nematode (B. lignicolus) in the United States. B. lignicolus also has been reported 
in France (1). 

With this first report, it was questioned as to how widespread the pine wilt 
nematode (B. lignicolus) is in the United States. Also, it was questioned whether 
the pine wilt nematode (B. lignicolus) is in the United States. Also, it was questioned 
whether the pine wilt nematode (B. lignicolus) was introduced into the United 
States from Japan or vice-versa. 

To help answer these questions, a survey of the state was started by the 
Department of Natural Resources, Division of Forestry to determine the host 
range and distribution of the pine wilt nematode (B. lignicolus). This paper 
describes the survey and reports the results to date. 

Methods and Materials 

Foresters in the Division of Forestry were asked to conduct the survey. They 
were given a training program and hand book on how to conduct the survey. When 
a forester observed a dying or dead pine tree, he would take increment core, wood 
chips, or branch sample and submit the sample to the forest pest specialist for 
determination of the pine wilt nematode (B. lignicolus). Information collected by 
the forester on the sampled tree included, county, township, range, section, species 
of tree, age, DBH, height, location, description of site and adjacent trees, presence 
of bark beetles in the tree and submission of specimens, if possible, estimate of time 
since tree death, and any stress factors on the tree. The Division of Entomology 
also conducted the survey through their field entomologists. The entomologists 
submitted samples and similar information as did the foresters to the forest pest 
specialist or to the state entomologist. 

Samples submitted to the forest specialist and state entomologist were soaked 

254 



Entomology 



255 



in water for 12 to 24 hours to extract nematodes. Nematodes collected were sub- 
mitted to the U.S.D.A., Animal and Plant Health Inspection Service for identifica- 
tion and confirmation of the pine wilt nematode (B. lignicolus). 

Results of the survey are reported to the foresters, entomologists, and U.S. 
Forest Service. 

Results 

The pine wilt nematode (B. lignicolus) has been identified, to date, from 23 
counties and confirmed by a nematologist from 14 of the 23 counties (Table 1). The 
survey to date has received samples from 48 counties. Eight pine species, Northern 
white cedar (Thuja occidentalis) and Eastern hemlock (Tsuga canadensis) have 
been sampled. Eastern white pine (P. strobus) is the only pine of the eight species 
not having pine wilt nematode (B. lignicolus) identified from it in Indiana (Table 1). 
The survey has sampled 185 trees from the 48 counties. The number of trees sampled 
by species are follows: pines - scotch (P. sylvestris), 114; red (P. resinosa), 25; 
eastern white (P. strobus), 6; Virginia (P. virginiana), 3; mugho (P. mugo), 4; 
shortleaf (P. echinata), 2; jack (P. banksiana), 11; Austrian (P. nigra), 7; and Pinus 
species, 11. Northern white cedar (T. occidentalis), 1. eastern hemlock (T. canaden- 
sis), 1. 



Table 1: Distribution of Pine wilt nematode, Bursaphelenchus lignicolus, in 
Indiana by county and host as of October, 1980. 



COUNTY 

Boone 1 

Clark 

Fulton 

Grant' 

Hamilton 1 

Harrison 

Hendricks 1 

Jasper 1 

Jennings 

Johnson 1 

Lake 

Lawrence 

Marion 

Martin 

Orange 

Perry 

Pike 

Porter 

Pulaski 

Putnam 1 

St. Joseph 1 

Shelby 1 

Tippecanoe 



HOST 

Pinus mugo 

P. spp. 

P. sylvestris 

P. sylvestris 

P. sylvestris 

P. sylvestris, P. virginiana 

P. sylvestris 

P. banksiana 

P. sylvestris, P. echinata, P. banksiana 

P. sylvestris, P. resinosa 

P. resinosa 

P. sylvestris 

P. nigra, P. sylvestris 

P. sylvestris 

P. virginiana, P. resinosa 

P. sylvestris 

P. sylvestris 

P. sylvestris 

P. sylvestris 

P. sylvestris 

P. sylvestris 

P. resinosa 

P. sylvestris 



'The nematodes found are identified as pine wilt nematode, but not confirmed by a nematologist. 



Of these trees sampled, the number identified with pine wilt nematode (B. 
lignicolus) are as follows: pines - scotch (P. sylvestris), 24 (15 confirmed); red (P. 
resinosa), 4 (2 confirmed); Virginia (P. virginiana) (1 confirmed); mugho (P. mugo\ 1 
(not confirmed); shortleaf (P. echinata), 1 (1 confirmed); jack (P. banksiana), 2 (1 con 
firmed); Austrian IP. nigra), 1 (1 confirmed); and Pinus species, 1 (1 confirmed). 



256 Indiana Academy of Science 

An additional 30 samples have nematodes collected from them, but not iden- 
tified at this time. 

No collection of pine wilt nematode (B. lignicolus) from an insect has been 
made. Nematodes were collected from under the elytra of a pine engraver (Ips spp.) 
but they were not the pine wilt nematode (B. lignicolus). Ips bark beetles, ceram- 
bycid larvae, and weevil larvae were reported from 38.9 percent of the trees sampled 
(Table 2). 25.7 percent of the samples having pine wilt nematode (B. lignicolus) had 
one of the above insects infesting the tree. Excluding the 30 samples the samples 
without pine wilt nematode (B. lignicolus) had 25 percent with one of the above in- 
sects infesting the tree. 12.8 percent of the samples with another nematode species 
present had one of the above insects infesting the tree. 12.1 percent of the samples 
without any nematodes had one of the above insects infesting the tree. 70 percent 
of the 30 samples to be identified have one of the above insects infesting the tree. 

Table 2: Number and percent of samples with or without bark beetles by 
presence or absence of nematodes. 





P WN 




Other Nematodes 


No Nematodes 


Total 


Bark beetles 


Positive 


Present 




Present 








Yes 


# 


% 


# 


% 


# 


% 


# 


% 


Yes 


9 


25.7 


41 


63.1 


22 


25.9 


72 


38.9 


No 


8 


22.8 


16 


24.6 


35 


41.1 


59 


31.9 


No Report 


18 


51.4 


8 


12.3 


28 


15.1 


54 


29.2 


Total 


35 


100.0 
18.9 


65 


100.0 
35.1 


85 


100.0 
45.9 


185 


100.0 
100.0 



No significant stress factors were reported for the trees sampled. Trees 
sampled ranged from one year dead to alive and averaged 6 months dead. No time 
after death was common for recovery of nematodes or the pine wilt nematode (B. 
lignicolus). Three trees with pine wilt nematode (B. lignicolus) were alive when 
sampled with the other trees ranging from 1 week to 1 year dead. These three 
trees did not have bark beetles attacking them. However, two of the trees were 
scotch pine (P. sylvestris) and found to be infested by Northern pine weevil 
(Pissodes approximatus). Trees not identified with the pine wilt nematode (B. 
lignicolus) ranged from alive to one year dead when sampled. 

Sampled trees ranged in age from 6 to 120 years and averaged 27 years. 
Diameter ranged from 2 to 22 inches and averaged 8 inches. Height ranged from 3 
to 80 feet and averaged 42 feet. 

Discussion 

The pine wilt nematode (B. lignicolus) is widespread in Indiana as indicated by 
being reported from 23 counties and seven pine species. The pine wilt nematode (B. 
lignicolus) is expected to be found in all Indiana counties and pine species eventually. 
The pine wilt nematode (B. lignicolus) is also widespread in the United States 
(Table 3) on seventeen pine species (Tabe 4) (4). This information helps to answer 
the questions of how widespread the nematode (B. lignicolus) is. Apparently the 
nematode IB. lignicolus) has been overlooked in the past because of lack cf 
knowledge regarding it. The question still remains to be answered whether the 
nematode IB. lignicolus) came from Japan or was introduced into Japan. 

The survey has not found any vectors; although the southern pine sawyer 
(Monochamus titillator) has been proven to be a vector in Iowa (5). The Ips bark 



Entomology 



257 



Table 3: 



Distribution of Pine wilt nematode, Bursaphelenchus lignicolus, in the 
United States -October, 1980 2 . 



Alabama 

Arkansas 

California 

Florida 

Illinois 

Indiana 

Iowa 

Kansas 

Kentucky 

Louisiana 

Maryland 

Michigan 

Minnesota 

Mississippi 



Missouri 

Nebraska 

New York 

North Carolina 

Ohio 

Oklahoma 

Pennsylvania 

South Carolina 

Tennessee 

Texas 

Vermont 

Virginia 

West Virginia 

Wisconsin 



Tersonal Communication, Kathryn Robbins, Pathologist, U.S., Forest Service, St. Paul, Minnesota. 

beetles may be vectors but it is unlikely as they do not enter the wood where it 
would be easier to contract the nematode (B. lignicolus). More insects need to be 
examined than have been examined by the survey to date for possible vectors. 

The Ips bark beetles reported in sampled trees may be the actual killers of the 
trees. The summer of 1980 was hot and dry in Indiana which encourages a build-up 
of the Ips bark beetles. 18.9 percent of the trees sampled were found with pine wilt 
nematode (B. lingnicolus) and 25.7 percent of these trees were infested with Ips 
bark beetles. 34 percent of the trees without pine wilt nematode (B. lignicolus) 
were infested with Ips bark beetles. Thus, 52.9 percent of the trees had a determined 
cause of death with 14.0 percent attributed to pine wilt nematode (B. lignicolus), 4.9 
percent to Ips bark beetles and pine wilt nematode {B. lignicolus), and 34 percent to 
Ips bark beetles. Also, the percent of those infested with Ips bark beetles increased 
in the late summer to 70 percent (26% of the last 37 trees sampled). Thus, Ips bark 
beetles may be the primary cause of mortality; but the relationship of the pine wilt 
nematode (B. lignicolus) and Ips bark beetles needs to be examined further. 



Table 4: 



Hosts of Pine wilt nematode, Bursaphelenchus lignicolus, in the United 
States - October, 1980\ 



Jack pine 

Swiss Stone pine 

Sand pine 

Sierra Nevada lodgepole pine 

Shortleaf pine 

Slash pine 

Mugho pine 

Austrian pine 

Longleaf pine 

Ponderosa pine 

Monterey pine 

Red pine 

Eastern white pine 

Scotch pine 

Loblolly pine 

Japanese black pine 

Virginia pine 



Pinus banksiana 

Pinus cembra 

Pinus clausa 

Pinus contorta var. murrayana 

Pinus echinata 

Pinus elliottii 

Pinus mugo 

Pinus nigra 

Pinus palustris 

Pinus ponderosa 

Pinus radiata 

Pinus resinosa 

Pinus strobus 

Pinus sylvestris 

Pinus taeda 

Pinus thunbergiana 

Pinus virginiana 



Tersonal communication, Kathryn Robbins, Pathologist, U.S. Forest Service, St. Paul, Minnesota. 



258 Indiana Academy of Science 

Literature Cited 

1. Foudin, A. and V. Dropkin. 1979. Pinewood nematode (Busaphelenchus 
lignicolus Mamiya and Kiyohara) - Missouri. Coop. PI. Pest Rep. 4:207. 

2. Mamiya, Y. 1972. Pinewood nematode, Bursaphelenchus lignicolus Mamiya 
and Kiyohara, as a causal agent of pine wilting disease. Rev. of PI. Prot. Res., 
5: 46-60. 

3. Mamiya, Y. and N. Enda. 1972. Transmission of Bursaphelenchus lignicolus 
(Nematode: Apelenchoididae) by Monochamus alternatus (Coleoptera: Ceram- 
bycidae). Nematologica 18: 159-162. 

4. ROBBINS, K. 1980. Personnal communication. 

5. Williams, D. J. 1980. Pinewood nematode (Bursaphelenchus lignicolus) Iowa. 
Coop. PI. Pest Rep. 5: 627. 



Insects and Other Arthropods of Economic Importance in Indiana During 1980 

Robert W. Meyer 

Department of Entomology 

Purdue University, West Lafayette, Indiana 47907 

Introduction 

The winter of 1979-1980 was relatively mild. Abundant snow cover (particularly 
in the southern part of the state) kept soils from freezing to their normal depth. A 
wet and cool March was followed by April, May and June rain deficits which 
gradually depleted the moisture in the saturated soils. The first 2 weeks of June 
were warmer than normal, but the very warm period in July was probably the 
most crucial for the corn crop. The heat combined with a shortage of moisture at 
pollination and silking time caused a number of anomalies that resulted in reduced 
fertilization and subsequent yield losses. Soybeans, which have a longer flowering 
period, suffered less than corn; soybeans were nevertheless shorter, the stems 
with fewer pods than normal at the base. The shorter and therefore more erect 
plants often failed to close the rows, encouraging weed growth. The drought 
stresses were most severe in the WC (west central) and NW districts, both impor- 
tant corn growing areas. On the other hand, the weather was excellent for planting 
and harvesting, both of which were completed ahead of normal. 

Corn and Small Grains 

Soil treatments were applied to about 47% of the 6,100,000 acres of grain corn 
in Indiana to prevent injury primarily by the corn rootworm complex (Western 
corn rootworm - Diabrotica virgifera, and northern corn rootworm - D. longicomis) 
in 1978, the last year for which firm data exist (5). Except for increases due to infla- 
tion, 1980 pesticide costs are not expected to differ a great deal. Two other factors, 
however, must be added. One involves pesticide efficacy: there were reports of in- 
secticide failures. The other involves the spread of economic populations into corn 
growing areas not previously seriously infested; to some extent the land once 
treated "for insurance," i.e., treated whether populations were known to be pre- 
sent or not, is becoming land where treatment (or crop rotation) is needed. In short 
where once only 40% of the treated acreage was actually infested, this year 
perhaps 50% or more was at risk. 

Surveys conducted during the last half of July and early August indicated a 
state average of beetles/plant double that of 1979, a jump from 0.30 to 0.63 
beetles/plant. The counties with the highest averages were in the EC district 
— Wayne, Union and Fayette counties averaged 2.0 beetles/plant. This is an exten- 
sion south and east of beetle stress. On a district basis, the NC and EC averaged 1.2 
the N W, WC and C averaged 0.9 adults/plant. Southern populations were all under 
0.2, and even the NE averaged only 0.4. Over the years the western corn rootworm 
(WCR) has gradually come to outnumber the NCR and this year WCR:NCR rela- 
tionships were 95:5 over all except in the southern districts. The two species were 
in about equal numbers in the SC; in the other southern districts the NCR still out- 
numbers the WCR. Although there undoubtedly were fields in which silks were 
damaged seriously by adults, especially where late-planted fields were near early- 
planted, infested fields, no such field was observed among the more than 400 
visited; however a rough, preliminary estimate puts at 70,000 the acres treated to 
protect silks. 

259 



260 Indiana Academy of Science 

Development proceeded as follows. First instar larvae were collected on 5 
June and on 18 June, again in Tippecanoe Co., 54% of the larvae were 1st instars, 
35% second and 11% third. A week later the seconds were the most abundant 
(47%), and on 2 July, when 46% were thirds, the first adult WCR was dug from the 
ground. Peak emergence from the ground of adults was reached during the week 
ending 23 July, for both species, and stick-trap catches peaked during the 7 days 
following. 

Adult western corn rootworms were collected for the first time from Brown, 
Floyd, Harrison, Crawford, Ohio, Switzerland and Clark counties. 

European corn borer (Ostrinia nubilalis) numbers fell to a state average of 68 
live 4th and 5th instar larvae/100 stalks as counted during the fall survey (Sept. and 
Oct.), near the 20 year (1961-1980) average of 62. Only in the northern districts were 
the averages above the 20 year levels. Starke Co. averaged 377/100 stalks, with 
numbers tapering off in all directions from that county. A new pupa was seen in 
Rush Co. on 20 May, but pupae were probably present earlier as 1st and 2nd instar 
larvae were collected in Knox and Gibson counties by 6 June. Adults were reported 
from black light traps on 28 May in Hendricks and LaGrange Cos. BL trap numbers 
peaked for the 1st time in the latter county on 11 June. There were no breaks be- 
tween the 1st and 2nd flights, or for that matter, between the 2nd and 3rd. Other 
peaks occurred on 13 August and 17 September. If a third generation occurs in In- 
diana it usually is confined to the SW district, but this year early instars were pre- 
sent at the end of September even in the northern districts. 

Attacks by the black cutworm (Agrotis ipsilon), the dingy cutworm (Feltia 
ducens) and sandhill cutworm (Euxoa detersa) on corn were reported but were 
minor problems, fewer than usual. Some wireworm reports, usually on corn in a 
wheat, soybean, soybean and corn sequence, were received, as well as a few army- 
worm {Pseudaletia unipuncta) - the latter in one case in a no-till field - cases. Most 
unusual was the loss of about 20% of a 126 acre field due to common stalk borer 
[Papaipema nebris). Normally active only at field edges, in this instance the larvae 
occupied large patches in a corn field planted no-till in rye. 

Corn leaf aphid (Rhopalosiphum maidis) infestation rates dropped from 47% 
infested in 1979 to 30% in 1980. These figures fail to bring out an unusual situation 
that is reflected by the jump in the rate of severely infested plants from about 1% 
in 1979 to 3% in 1980. About mid-July, populations in excess of 20,000/plant began 
to appear on grain corn particularly in the WC district. They covered not only the 
tassel (in fact, because that portion suffered an early decline as a result of high 
temperatures and drought, it was not always infested) but also the top leaves (not 
necessarily the topmost); such colonies were highly visible, instigating many 
grower inquiries. Often these infested leaves were intermingled with whitened, 
blasted leaves, heightening the contrast. These infestations appeared first in fields 
planted in sandy or gravelly soils near the Illinois line, and on hill tops, although 
not all stressed fields were infested with aphids. Undoubtedly such numbers added 
to stresses already present, but most of the loss was probably due to the drought. 
Five percent of the plants surveyed were barren, and another 5% produced mere 
nubbins. In more normal years only 1% or less fall into each of these categories. 

In that connection, smut galls were more common than usual; 6% of the plants 
were infected, double the infection rate of last year. The counties from Clinton to 
Grant and as far north as Miami were the most heavily infested, averaging up to 
21%. Weed control, on the other hand, was generally good. 

Only 2% of the ears showed any signs of bird attack, and a similar number 



Entomology 261 

showed feeding by the corn earworm (Heliothis zeae) and the fall armyworm 
(Spodoptera frugiperda). Of the larvae still present at survey time, 89% were corn 
earworms, 11% fall armyworms. Because of the poor fill at the ear tips, kernel 
losses to these larvae were minimal. 

The oat bird-cherry aphid {Rhopalosiphum padi) was present in only 13% of 
the corn fields, just half those of last year, and they were rarely numerous. 

The mean percentage infestation by the Hessian fly (Mayetiola destructor) for 
wheat was 1%, very nearly the low figure that applied in 1979. There was a mean 
of 1.2 puparia/100 stems. 

Forage Legumes and Soybeans 

An estimated 200,000 acres of soybeans were treated for the control of Mex- 
ican bean beetles (Epilachna varivestis) during 1980. Beetle attack reached its 
highest levels in Owen and Clay counties with extensions in contiguous counties as 
far south as Daviess Co., and westward into Vigo Co.; Monroe Co. had heavier in- 
festations than usual, and Morgan, lighter. Elsewhere only scattered fields reached 
economic levels. The beetle tends to extend its soybean territory northward each 
year. Two years ago Richmond on the east side of the state marked its most nor- 
thern extension. This year a heavily infested field was seen on the outskirts of Win- 
chester 25 miles north. For the second year a light infestation was reported from 
Tippecanoe Co. In summary, while the beetles were probably more numerous this 
year, and more widespread, the damage was less, and probably only half the 
treated acreage was profitably treated. 

Adults were reported as early as 3 May from Franklin and Jackson Co. alfalfa 
fields, and from trap crop snap beans in Jennings Co. on 21 May. Egg masses were 
present on soybeans in Monroe Co. by 4 June and 1st instars by 10 June. The 
following data were collected at 2 week intervals in 10 soybean fields in Bar- 
tholomew and Jennings counties by sweeping. Overwintering adults averaged 
0.5/sweep on 17 June, 1st generation adults, 2.7/sweep on 27 July. First generation 
larvae of all ages averaged 0.9/sweep on 22 August, with 3rd and 4th instars 9 
times as common as early instars. 

An estimated 20,000 acres of soybeans were treated for the control of green 
cloverworm (Plathypena scabra), especially when grasshoppers (Melanoplus spp.) 
added to leaf loss. The latter were more common this year than usual, and often in- 
flicted conspicuous damage especially at field edges. 

Because of the scattered nature of the attack, it is difficult to estimate the 
number of of acres that were attacked by bean leaf beetle (Cerotoma trifurcata). 
This insect, which normally ends its activity when the beans enter their reproduc- 
tive phases or before, this year attained economic numbers in soybeans at pod-fill 
stages as late as August. Most of these attacks occurred in Sullivan and Daviess 
counties. 

A leaf-mining beetle, Odontota horni, was observed at the rate of about 1/sq. 
meter in a soybean field in Starke Co. These adults had probably emerged from 
soybean leaves as abandoned mines were present. Adults of this species were 
numerous enough to be noted from beetle samples collected from the shores of 
Lake Michigan. The species, in the same genus as the locust leaf minor (O. dorsalis) 
which more frequently attacks soybeans, is said to breed normally in tick-clover 
(Desmodium rigida). The larvae have not been described. Isolated specimens of 
this species have been collected from soybeans before, but never in such abun- 
dance. 



262 Indiana Academy of Science 

The soybean cyst nematode (Heterodera glycines) was reported for the first 
time from Benton Co., a new county record. They were collected from the roots of 
soybeans 2 September near Earl Park by John Ferris who also identified them. 

An estimated 25,000 acres of alfalfa were treated for the control of alfalfa 
weevil (Hypera postica) larvae in alfalfa; 20,000 of these were in the state south of 
US 50, and the remainder between that highway and Indianapolis. Treatment 
elsewhere was sporadic. Treatment south of Indianapolis was probably inadequate 
as severe defoliation was observed as far north as Vermillion Co. on the west side 
of the state and Randolph on the east. Many growers were able to use the early 
harvest of the first cutting as a control measure. In less severe attacks, regrowth 
almost masked the damage done by larvae at their peak numbers; this situation 
was common. 

Three hundred thirty alfalfa samples 15 cm 2 from throughout the state col- 
lected during November and December of 1979 averaged 5.6 eggs/sample with no 
significant differences between the northern, central or southern districts (Range 
in means from to 16.9). (In the fall of 1978 the southern districts had 4 times as 
many eggs as the northern, the average was 17 and the range 1-74). By the end of 
April, 1980, larvae averaged 4/infested stem in alfalfa 26cm tall in the SC district, 
averaged 84% infested. At the same time WC infestations averaged 22%, with 0.7 
larvae/infested stem in alfalfa 16 cm tall (Infestations ranged from 0-72%). In mid- 
May northern districts ranged from 0-42% infested (average 20%) in alfalfa 30 cm 
tall, with about 1.5 larvae/infested stem. 

Probably fewer than 50,000 acres of alfalfa were treated for the control of the 
potato leaf hopper (Empoascafabae) during 1980. The species was less of a problem 
than normal. 

Vegetables 

(This vegetable summary, provided by Alan C. York, was based on numerous 
visits to both commercial and private acreages, and telephone and written reports 
received through the extension services.) Only the imported cabbageworm {Pieris 
rapae) appeared in normal - and damaging - numbers on cole crops. Cabbage looper 
(Trichoplusia ni) and diamondback moth {Plutella xylostella) larvae were fewer 
than normal, there were a few reports of zebra caterpillar (Ceramica picta), and 
cabbage aphids {Brevicoryne brassicae) were virtually absent. 

Colorado potato beetles (Leptinotarsa decemlineata) were in below-normal 
numbers in both generations with the possible exception of central Indiana. Most 
other pests of solanaceous crops were at normal levels: potato leafhopper {Em- 
poasca fabae), potato flea beetle (Epitrix cucumeris), and tomato fruitworm 
(Heliothis zea) (there was some influx of those from early maturing field corn). 
Aphids, both the green peach (Myzus persicae) and the potato (Macrosipkum 
euphorbiae), were almost absent. The tobacco horn worm (Manduca sexta) on the 
other hand was more numerous than at any other time in the last six years, at least 
in the NW district. Some tomato fields there were stripped of leaves from early to 
mid-September. 

European corn borer (Ostrinia nubilalis) larvae attacking several categories of 
garden produce, were extremely numerous in September in what appears to have 
been a third generation. 

Squash vine borer (Melittia satyriniformis) larvae were early and numerous 
again, and the squash bug (Anasa tristis) was more numerous. Striped cucumber 
beetle (Acalymma vittatum) adult activity was brief and low in number the first 



Entomology 263 

generation. The second generation was normal. Spotted cucumber beetle 
(Diabrotica undecim punctata Howard) numbers were also about average. 

The aphid Brachycolus asparagi was collected for the first time in the state by 
James Clark, 27 August, a new state record. It was subsequently collected in 
Johnson Co. (8 Sep. by John Favinger), Vigo Co. (Terre Haute) and Clay Co. (Brazil) 
by Earl Huff and Virgil Knapp 21 Oct., Hendricks Co. (North Salem, 21 Oct.) and 
Madison Co. (Anderson, 22 Oct.) by Virgil Knapp. All determinations, by Virgil 
Knapp, were confirmed by Manya B. Stoetzel, USDA. These are, of course, all new 
county records for this aphid, which attacks asparagus ferns, causing conspicuous 
witches brooms in addition to the usual aphid damage. 

Ornamentals, Forest, Shade and Fruit Trees 

The Japanese beetle (Popillia japonica) was collected from the following coun- 
ties by means of lure traps this year - all new county records. (County names are 
followed by the name of the nearest town, the date the trap was recovered and the 
name of the collector). Floyd (Greenville 15 Aug., Mark Bratovich), Vermillion (Clin- 
ton 15 Aug., Earl Huff), Johnson (Greenwood 15 Aug., Rudy Carandang), Jay 
(Portland 17 Aug., Harry Bollinger), Gibson (Johnson 18 Aug., Mark Bradovich), 
Owen (Vandalia 20 Aug., Robert Heaton), Madison (Anderson 4 Sept., Virgil 
Knapp), Boone (Zionsville 7 Sept., Virgil Knapp), Hamilton (Noblesville 9 Sept., 
Virgil Knapp) and Carroll (Delphi 17 Sept., Virgil Knapp). Determinations by the 
collectors were confirmed by James Clark. 

The gypsy moth (Lymantria dispar) was reported earlier this year from Vigo, 
Hendricks, Wayne and Franklin counties elsewhere (4). In addition to those new 
county recoveries, the following are added here: from Elkhart and Goshen in 
Elkhart county by W. Felts on 12 Aug.; from Ellison in Allen Co. by Harry Boll- 
inger on 8 Sept. Determinations by the collectors were confirmed by James Clark. 

Larvae near pupation of the spruce budworm (Choristoneura fumiferana) 
were collected from blue and Norway spruce in Putnam Co. about 6 June by Philip 
Marshall, and eggs and empty pupal cases were collected by Cedric Durkes in 
Lafayette (Tippecanoe Co.) from spruce 11 June. Determination of the first was by 
Philip Marshall, the Tippecanoe Co. record by Donald L. Schuder. These are both 
new county records, and only the 2nd and 3rd time that this species has been 
observed overwintering in this state. (An earlier record was from Lake Co.) Large 
numbers of adults entered the state in 1974; unless these are the result of that inva- 
sion, even such a large invasion was unsuccessful. 

Linden looper (Erannia tiliaria) and halfwing geometer (Phigalia titea) larvae 
were again rather common on SC trees by early May. They caused less damage this 
year than last. Eastern tent caterpillar [Malacosoma americanum) was also rather 
common in Washington and Harrison as well as Pike counties. Cottony maple scale 
{Pulvinaria innumerabilis) was again very common on urban maples in a number of 
larger cities. While they attracted a great deal of attention, the oystershell scale 
(Lepidosaphes ulmi), which was also very common, escaped attention. 

The pinewood nematode (Bursaphelenchus lignicolus) was reported from 
Porter, Tippecanoe, Marion, Clark (1), Hendricks (2), Perry, Martin, Pike and Jenn- 
ings (3) counties this year, all new county records. In addition since those reports 
the species has been collected from the following counties reported here for the 
first time. All were from Scotch pine except the Lake Co. record, which was from 
red pine. (The county name is followed by the collector's name and the nearest 
city.) Marion (Philip Marshall, Indianapolis), Lawrence (D. Ernest, Mitchell), Fulton 



264 Indiana Academy of Science 

(Philip Marshall, Rochester) Pulaski (R. B. Cummings, Winanac) and Lake (R. B. 
Cummings, Winanac). (Determinations by A. Foudin, USDA.) 

Five pheromone traps in Vincennes caught a total of 2900 redbanded leafroller 
{Argyrotaenia velutinana) males during the trapping season, the highest level in 5 
years and approached only once - last year (2300). Peak catch was taken during the 
last 2 weeks of April. The Tippecanoe Co. peak catch occurred during the 1st week 
of May; Tippecanoe Co. totals were also the highest in 4 years. Codling moth 
[Laspeyresia pomonella) numbers reached a 4 year high in the latter county. 
Numbers of all other apple and peach moths were not significantly different from 
other years in either county except for a great drop in the numbers of lesser 
peachtree borers (Sanninoidea exitiosa) from 7200 last year to 1500 this year, in 
Knox Co. 

The aphid Hyadaphis tataricae was collected for the first time in the state on 
22 Sept. by Bruce Cummings - a new state record. It was collected from Lonicera 
tatarica, a variety of honeysuckle, in Rensselaer, in Jasper Co., a new county 
record as well. Determination was by Virgil R. Knapp, confirmation by Manya B. 
Stoetzel, USDA. 

Man and Animals 

There were only 5 confirmed cases of St. Louis encephalitis in the state, all 
from Knox Co., by 1 Nov, and 2 cases of La Crosse strain — 1 each from Lake and 
Morgan counties. 

Beneficial Organisms 

The ratios (in percent of the total of these species) among the spotted (Col- 
eomegilla maculata), the convergent (Hippodamia convergens), the 13-spotted (H. 
tredecimpunctata) and Cycloneda sanguinea as indicated by sticky trap collections 
in a corn field in Tippecanoe Co. in 1977, 1978, 1979 and 1980 follow: 27:65:3:5; 
69:6:16:9; 81:9:3:7 and 58:7:2:33. For the first time in the 4 seasons of trapping, 
sanguinea has exceeded the single-digit range. Because of the small numbers of 
this species collected in past years, it is difficult to state where peak activity is, and 
to decide whether a major outbreak of corn leaf aphids was a factor in the increase. 
The numbers of both Coleomegilla maculata and Cycloneda sanguinea increased 
following the mid-July burst of aphid activity, but in the case of the former, the in- 
crease at that time occurs every year even without aphid explosions. The traps 
were in the same field every year, but for the first time the whole field was treated, 
including the trap area, for corn rootworms. The total of coccinellids collected has 
increased each year, but there was a drop of about a fourth in Coleomegilla 
numbers this year. 

About 28% of 297 alfalfa weevil (Hypera postica) adults collected in the first 3 
weeks of May were parasitized by the braconid Microctonus aethiopoides. Weevil 
collections were made primarily in the central districts (224 of 297). Reported 
earlier this year were the new county records from Fulton, Boone, Decatur, Henry, 
Monroe and Lawrence counties (3). Reported here for the first time are the follow- 
ing new county records: (nearest city and date in parentheses) Fountain (Rob Roy 
19 May), Rush (Homer 6 May), Delaware (Royerton 12 May), Randolph (Farmland 
12 May), Union (Liberty 6 May) and Jennings (San Jacinto 21 May). All were reared 
from weevils by the author whose determinations were confirmed by Arwin Pro- 
vonsha. 

About 27% of 880 alfalfa weevil larvae collected the last week of April were 



Entomology 265 

parasitized by Bathyplectes spp. Ninety percent of the larvae were collected from 
the SW and SC districts, and 88% of the parasites were B. anurus. This species was 
collected for the first time from Henry, LaPorte, Vermillion, Sullivan and Jackson 
counties. The Vermillion, LaPorte and Henry records are the first from north of In- 
dianapolis. 

Literature Cited 

1. Anon. 1980. USDA. APHIS, Coop. Plant Pest Rep. 5(16):307. 

2. 1980. USDA. APHIS. Coop. Plant Pest Rep. 5(17):328. 

3. 1980. USDA. APHIS. Coop. Plant Pest Rep. 5(30):562,564. 

4. 1980. USDA. APHIS. Coop. Plant Pest Rep. 5(34):645. 

5. Park, Earl. 1980. Pesticide Usage. Annual Crop and Livestock Summary 
1979:67-69. 



Effect of Photoperiod and Temperature upon the Growth Rate of the Later Instars 
of Erythemis simplicicollis (Say) (Odonata; Libellulidae) 

B. El wood Montgomery 

Purdue University, 

West Lafayette, Indiana 47907 

During the period of 1961 to 1968 over 9,000 Odonata naiads were collected in 
the field and introduced into photoperiod-temperature studies in our laboratory. 
Records of several experiments have been published (6, 7, 9, 10, 11, 12). However, 
many other experiments have never been analyzed and published because of pre- 
occupation with studies in other areas. An attempt is being made now to sum- 
marize and analyze these and to publish results that seem significant. 

Erythemis simplicicollis (Say), sometimes called the "green jacket," is a very 
common libellulid dragonfly of wide distribution. It occurs from southern Canada 
into Mexico and the Antilles. In Indiana it is virtually ubiquitous around ponds and 
lakes. It has a flight period in the state from early May into October, being most 
abundant from mid-June to late August (8) (Table 1.). Bick (1) from published data 
by Davis and Fluno (2) and his own observations gave the flight period in Florida 
and the Gulf States from March 4 to November 2, and noted oviposition "con- 
tinuous" from April 10 to September 4. In more southern areas it may be found in 
flight throughout the year. 

The life history of this species at New Orleans was worked out by Bick (1) who 
reported an incubation period of 10-16 days, with an average of 11.6 days, and a 
duration of 113 days for the one individual which survived to emergence and an 
average of 109.3 days for the 11 individuals in his study (Table 2). No conditions of 
temperature or day length were cited, except that the eggs were placed "under a 
bell jar above which a desk lamp was kept burning throughout the incubation 
period." The rearing apparently was carried out under "room conditions" and 

Table. 1 Relative seasonal abundance of adults and different instars of 
Erythimis simplicicollis (Say). 





April 


May 


June 


July 


Aug. 


Sept. 


Oct. 


Index of 
adult abun- 
dance 1 




-1-5 


5-15-10 


16-15-23 


15-13 12 


10-2-1 




Number 


73 


47 






380 


165 


45 


Octult 










2% 






Septult 










8% 




2% 


Septult 










21% 


1% 


4% 


Quintult 




2% 






29% 


8% 


13% 


Quintult 


4% 


6% 






26% 


14% 


24% 


Tertult 


33% 


13% 






11% 


19% 


33% 


Penult 


33% 


.30% 






4% 


35% 


13% 


Ult 


30% 


49% 






2% 


23% 


9% 



'Based upon data from Montgomery (1945); the index of relative abundance was compiled by tabulating the 
number of collections, or sightings, over a 40 year period, by thirds of months. 

^ased upon data from Montgomery (1971); compiled from analysis of collections of maiads made for 
temperature-photoperiod studies. 



266 



Entomology 



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268 Indiana Academy of Science 

natural day length. The natural day length at New Orleans (30°N latitude) at the 
time of hatching (June 18-24) was 14 hours 4 minutes, the longest days of the year 
at this latitude. This gradually decreased (ca Vz to 2 minutes per day) to 11 hours 40 
minutes on October 9, the date of the emergence of the naiad which completed 
development. 

A collection of naiads, in the nontult to quintult instars, was obtained August 

9 from a lake in an old strip mine pit at Ashboro, Clay County, Indiana. These were 
separated into individual culture dishes during the following two days and reared 
through the remaining instars in our laboratory. Ten naiads were placed in each of 
15 "climate" cabinets with different regimes of temperature and photoperiod. 
These were selected to measure the effects of temperature and photoperiod upon 
development, especially to determine if there is a definite division between "long 
day" and "short day" environments for this species, and to compare the length of 
time required for a complete life cycle with that reported by Bick (1) in Louisiana. 

The day length at the time the naiads were collected and removed from the 
natural environment (40° N latitude) was approximately 14 hours, decreasing 
about two minutes per day. 

A mated pair of E. simplicicollis was collected July 10, 1967 at a pond on the 
Purdue Forage Farm in Dubois County, Indiana. A clutch of eggs was obtained by 
grasping the female by the wings folded above the body and dipping the tip of the 
abdomen against the surface of water in a small container. The eggs were returned 
to the laboratory, which was air conditioned to 21 °C, and incubated under natural 
light conditions (day length of 14 hours, 50 minutes to 14 hours, 30 minutes). A 
number of eggs hatched in 14 days and naiads were isolated into 2" culture dishes. 
Lots of 10 were placed in cabinets at three different temperatures, 18°C, 21 °C and 
24 °C, all with a 16-hour photoperiod. Most of the naiads survived through the first 
instar, many through the second, a few through the fourth at 18°C, and the fifth at 
21 °C and 24 °C. The data from the 21 °C rearings were combined with those of the 
August 9 material of the same temperature to give a complete life history from the 
Indiana population. This furnishes a base for comparison with Bick's life history 
data based upon a Louisiana population (Table 2). 

Naiads hatched from eggs in the laboratory and those of the size of the 
Ashboro material (2-5 mm in length) were started in 2" culture dishes, transferred 
to 4-V4 " dishes when they became conspicuously larger (7-9 mm in length) and to 
1-or I-V2 pint fruit jars when the ultimate instar was attained. Inverted U-shaped 
strips of l k " screen wire were provided as supports for emergence. All dishes and 
jars were examined daily for molts and to supply food. It may be noted that those 
environments indicated as hours photoperiod were thus exposed to light for 5 to 

10 minutes daily for this examination. Food consisted of protozoa and other 
organisms from "hay infusions" and/or nematodes for small naiads and "white 
worms" (Enchytraeidae) for all others. The worms remained alive and active from 
one feeding to the next so that it is believed that food was always available. The in- 
star of naiads which did not survive to emergence was determined by the length of 
the wing pads of the dead insects, which were preserved with all exuvia until 
checked. However, this character could not be employed to identify the instar of 
naiads before the septult as the pads are not present until this instar and are too 
small to be reliable in this and the following instar. Furthermore, neither this nor 
any other instar. characteristic can be examined without danger of injury in living 
naiads. 

Upon the assumption that all naiads develop in 13 instars the octult of our 



Entomology 269 

material has been equated with the sixth instar of Bick's study. There appears to 
be some variation from the instar characteristics given by Bick but a more 
thorough study of our extensive material is needed to determine if there are dif- 
ferences between the two populations, or if the number of instars may vary be- 
tween individuals, as was found in Anax (6). The photoperiods in our studies were 
constant as the cabinets were light proof and all examinations were made during 
light periods. However, the length of the effective photoperiod in nature may be 
questioned as there appears to be no definitive evidence on the threshold of optical 
stimulation of Odonata naiads. There is a little experimental evidence that this 
threshold, in some species at least, may be below the level of moonlight (5). This 
series of experiments shows very definite effects of photoperiod as well as 
temperature upon the development, or the duration of instars (Tables 2-5). 

The survival rate of Odonata naiads in laboratory rearings is very low. Bick 
had one naiad of 11 reach emergence (9%) with two others reaching the ultimate in- 
star but dying after 1 and 15 days respectively. In most of our experiments, 
especially with Anisoptera, the rate of survival to adult emergence was about the 

Table 3. Duration of instars in several regimes of temperature/photoperiod. 













Instar 










Temperature/ 


Naiad 


















photoperiod 


Number 


6 


7 


8 


9 


10 


11 


12 


13 



3297 1 14 8 17 17 21 49 56 



27°/ 3288 8 7 10 21 20 



16 hrs 3295 11 12 43 37 52 33 



3291 



3292 8 28 17 19 70* 

3324 1 25 8 11 28 22 39 117 

3320 8 9 37 14 22 35 58 

3322 15 16 12 21 27 72 

3323 15 21 21 33* 



27°/ 3325 



13V 2 hrs 3326 15 16 12 19* 



23 14 28 209 



3319 15 32 



3321 11 23 28 35 134 



39 18 43* 



3376 12 58 23 15 16 21 70 115 

27°/ 3368 8 116 43 92 64 92 

9 hrs 3372 43 96 32 21 62 62 



3375 26 105 24 29 34 





3373 






108 


21 


52 


75 


80 




3359 


19 


7 


31 


67 


26 


25 


43 


27°/3x3 hrs 


3362 




46 


11 


10 


20 


30 


67 



3355 22 13 128 



27°/0 hrs 3357 19 143 71 37 



3348 19 120 55 101 64 

3349 25 102 49 77 59 





3354 








8 


121 


50 


71 


100 




3434 


30 


184 


42 


34 


142 


103 


50 


142 


21°/0 hrs 


3437 






24 


173 


39 


41 


121 


146 



270 



Indiana Academy of Science 



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272 Indiana Academy of Science 

same. However, in these experiments, probably because the tests were started 
with naiads instead of eggs, the rate is somewhat higher. The rate of survival to 
emergence in nature must be very low as the eggs produced per female are 
numbered in the hundreds and thousands (2). We counted 250 and 990, respective- 
ly, in two clutches of eggs in this species (13) and it appears likely that each female 
may oviposit several times. Jacobs (4) reported another libellulid, Plathemis lydia 
(Drury) as ovipositing seven times at the same pond in a period of 22 days; we found 
one clutch of P. lydia eggs to number 1429. 

The "climates" in which the lots of naiads of the August 9 collection were 
reared included nine photoperiods at 27°C (80°F), and three each at 21 °C (70°F) 
and 34°C (90°F). These were selected to measure the effects of temperature and 
photoperiod especially to determine if there is a definite division in photoperiod 
lengths upon the rate of development. At 34°C only three naiads survived beyond 
the initial instar (that in which collected), all of these were collected in the quintult, 
the remaining 27 were collected in the septult or younger rendering instar iden- 
tification impossible. All 10 naiads in the nine hour cabinet died without molting. 
Two quintult naiads in the 0-light cabinet survived into the tertult (in 57, 56, and 16 
days) and into the ult (18, 25, 17 and 16 days) respectively. The one in the 16-hour 
environment reached the adult in 2, 28, 17, 21 and 68 days. 

The data on development in the 21 °C and 27 °C environments, expressed in 
duration of instars (minimum, maximum and average) are given in Table 4. The 
average length of time required for completion of development through the last 
seven instars (7-13) varied from 124 to 236.2 days for the photoperiods of 13 hours 
and longer. However, this variation is not at all correlated with the length of 
photoperiod as the shortest period was at 13 hours and the longest at 13 Vz hours. 
In view of the variation of duration in all instars for individuals these differences 
are probably not significant. However, the 12-hour period appears to be less 
favorable, although the data are incomplete, as four died in the initial instar and 
two surviving into the ult both died after a few days. The 9-hour period showed a 
considerably longer time for this development but when the nine hours were divid- 
ed into three equal periods distributed at equal intervals throughout the day the 
time required was reduced. At 21 °C the period for completion of the life cycle was 
lengthened, although at 16 hours of light it was less than those of the 27°C/9- and 
3x3-hour periods. 

In all regimes the duration of instars was greater than the periods reported 
by Bick for Louisiana; the ratios for the averages as shown in Table 5, varied from 
0.9 (only two cases) to 36.0 times those in Bick's rearings. In all except the 27°C/13 
hour there were one or more naiads with three times or more instar lengths than 
the maximums of the Louisiana material. These "long" periods were found in all in- 
stars included in the experiment (5-13), and showed some correlation with 
temperature and photoperiod. The percentages of "long" periods of all naiads in all 
instars for the 27°C groups were: 16h.-18°/o, 14-V2h.-12°/o, 14h.-16°/o, 
13-V 2 h.-13°/o, 12h.-40%, 9h.-53°/o, 3x3h.-29°/o, and Oh. -52% and for 21°C 
groups: 16h.-21%, 9h.-59°/o and Oh. -93%. The effects of 16 hours of light at 
21°C and the 3x3 photoperiod at 27°C in decreasing the number of such long 
periods is evident. The ability of naiads of this species to complete development 
with only 5-10 minutes of light in each 24-hours, with repeated very long instars 
resulting in 6.4 and 8.0 times the period required under "natural" conditions is 
most interesting. The universal occurrence of these long periods must be an adap- 
tation to survival over winter. The very definite difference in the effects of the 
13-and 12-hour photoperiods correlates very well with the flight period in the In- 



Entomology 273 

diana season, mid-May through September. A day length of 13 hours or more, 
beginning about April 10 would seem to stimulate emergence of some individuals 
within a month and a day length of 12 hours and less beginning September 1 would 
tend to inhibit the emergence of most individuals until the combination of lower 
temperatures and shorter day lengths would end all emergences for the season. 

E. simplicicollis appears to have only one generation a year and to pass the 
winter in all instars as young as the octult, possibly younger as we have no data on 
either the "long" instars earlier or any occurrence of the earlier instars in the 
spring or autumn (Tables 1-3). There are no definite broods, ovipositing continuing 
through most of the flight period as observed by Bick (1), and development being 
lengthened or interrupted, by the lower temperatures and shorter day lengths of 
winter in any instar. However, it seems possible that a few individuals might com- 
plete the life cycle during a single season and that others might go through two 
winters. Furthermore, it is very likely that the shorter developmental period in the 
Louisiana population with a longer season of favorable growth may result in a par- 
tial second generation and that in more southern areas there could be two genera- 
tions a year. In the northern portion of the range with opposite conditions it is 
probable that more than a year is regularly required for a generation. 

Literature Cited 

1. BlCK, George H. 1941. Life-history of the dragonfly Erythemis simplicicollis 
(Say). Ann. Ent. Soc. Amer., 34:215-230. 

2. Corbet, Philip S. 1963. A biology of dragonflies. Chicago, Triangle Books, 
xvi + 247 pp. 

3. Davis, E. M. & J. A. Fluno. 1938. Odonata at Winter Park, Florida, Ent. 
News, 49:44-47. 

4. Jacobs, M. E. 1955. Studies on territorialism and sexual selection in 
dragonflies. Ecology, 36:566-586. 

5. Jenner, Charles E. & Paul E. Lutz. 1963. (Discussions during Colloquium on 
the Odonata.) Pr. N. centr. Br., ent. Soc. Amer., 18:101-156. 

6. Macklin, Jerry M. 1964. Notes on the life history of Anax Junius (Drury) 
(Odonata: Aeshnidae). Pr. Ind. Acad. Sci., 72:154-163. 

7. & B. Elwood Montgomery. 1962. Further notes on the rates of develop- 
ment of naiads of Neotetrum pulchellum (Drury) (Odonata: Libellulidae). Pr. 
Ind. Acad. Sci., 2:158-160. 

8. Montgomery, B. Elwood. 1945. The distribution and seasonal abundance of 
the Indiana species of Corduliidae and Libellulidae. Pr. Ind. Acad. Sci., 
54:217-224. 

9. 1963. Effects of photoperiod and temperature on rate of development in 

naiads of Ischnura verticalis (Say). Pr. N. centr. Br., ent. Soc. Amer., 
18:140-146. 

10. 1971. Records and observations of Indiana odonata. Pr. Ind. Acad. Sci., 

80:253-263. 

11. & Jerry M. Macklin. 1962. Rate of development of the later instars of 

Neotetrum pulchellum (Drury) (Odonata: Libellulidae). Pr. N. centr. Br., ent. 
Soc. Amer., 17:21-23. 

12. 1966. Photoperiod studies on the Odonata. Pr. N. centr. Br., ent. Soc. 

Amer., 21:30-31. 
13. Zehring, Connie Sue, Archie Alexander & B. Elwood Montgomery. 1962. 

Studies of the eggs of Odonata. Pr. Ind. Acad. Sci., 72:150-153. 



New Distribution Records for Mosquitoes (Diptera: Culicidae) in St. Joseph County, 

Indiana 

David B. Taylor 
Vector Biology Laboratory 
University of Notre Dame 
Notre Dame, Indiana 46556 

Introduction 

Prior to the formation of the St. Joseph County Mosquito Surveillance Pro- 
gram in 1976 only one species of mosquito, Aedes hendersoni, had been recorded 
from the county (4). During the first year of the Surveillance program over 2,000 
larval sites were sampled and more than 15,000 adults collected in 13 New Jersey 
Light Traps. As a result of this program 35 species of mosquitoes were reported 
from the county (5). Subsequent to 1976 the program has continued to monitor the 
mosquitoes of the county. 16 New Jersey Light Traps have collected more than 
300,000 adults, over 5000 larval sites have been identified and 4 new species record- 
ed (Table 1). This program has allowed us to determine what potential vector 
species of mosquitoes are present, to propose control measures and to monitor 
population size. 

Materials and Methods 

A series of 16 New Jersey Light Traps was placed throughout the county in 
many different habitats including urban, suburban and rural sites (Figure 1). The 
traps were operated 7 nights a week and collections were made 3 times a week 

Table 1. The mosquitoes of St. Joseph County, Indiana** 

Aedes Coquillettidia 

abserratus perturbans 

atropalpus (Restifo & Lanzaro 1980) Culex 

aurifer erraticus 

canadensis pipiens 

cinereus restuans 

dorsalis salinarius 

excrucians tarsalis 

fitchii territans 

flavescens* Culiseta 

hendersoni inornata 

sollicitans* melanura 

sticticus morsitans 

stimulans minnesotae 

triseriatus Orthopodomyai 

trivittatus alba 

vexans signifera 

Anopheles Psorophora 

barberi ciliata 

crucians columbiae 

punctipennis cyanescens* 

quadrimaculatus ferox 

walkeri Uranotaenia 

sapphirina 

•• Shroyer et al. 1977 
* New records 

274 



Entomology 



275 




Figure 1. Locations of light traps in St. Joseph County, Indiana. 



from the beginning of May until the middle of October for the summers of 1976 
through 1980. During the summers of 1976 through 1979 a larval survey was con- 
ducted by dividing the county into square mile sections and systematically inspect- 
ing each section twice a year for mosquito breeding sites. Several other 
surveillance techniques have also been used such as Fay, suction, and oviposition 
traps. Representative specimens have been placed in either the collection of the 
author or that of the University of Notre Dame. 

Results and Discussion 

During the summer of 1979 an intensive urban survey was conducted to locate 
the possible breeding sites of Ae. triseriatus, the vector of LaCrosse Encephalitis, 
in urban areas. During this survey larvae of Aedes atropalpus (Coquillett) were 
found in tires at two locations on the south side of South Bend (6) (Figure 2). Ae. 
atropalpus normally is found in rock pools along the shores of lakes or streams in 
northern and eastern United States. The strain of Ae. atropalpus found in tires, 
unlike its feral conspecifics, feeds readily on man; therefore, it could become a 
health threat as a domestic species (2). Despite an intensive search no Ae. 
atropalpus were found during the summer of 1980. This leaves doubt as to whether 
or not this species has become established in the county. 

On 22 September 1978 a single female of Psorophora cyanescens (Coquillett) 



276 



Indiana Academy of Science 




Figure 2. Sites of new species records in St. Joseph County, Indiana. 



was collected biting a human near St. Joseph Lake on the campus of the University 
of Notre Dame (Figure 2). Ps. cyanescens is primarily a species of the southeastern 
United States (1). It has been collected in 10 other counties in Indiana (Figure 3), 
the northernmost of which was Wayne County. The present record for St. Joseph 
County is the most northern record of this species in the United States. 

During the past 5 years 340,860 adult mosquitoes have been collected in the 
light traps. Table 2 gives a summary of the light trap data for the years 1976 
through 1980. The total number of mosquitoes has increased every year since the 
program began. Relocation of some of the traps and increasing the number from 13 
to 16 could be partially responsible for this increase. The magnitude, however, in- 
dicates that there has been an actual increase in the number of mosquitoes over the 
past five years. 

On 1 June 1979 a single female of Aedes flavescens (Mueller) was collected in 
the Lakeville light trap. No larval sites were found in the vicinity of the light trap 
and there have been no additional specimens collected. Siverley stated that this 
species is predominately a prairie species and has been recorded from only two 
other counties in Indiana, Delaware and Steuben (Figure 3). 

On the 14th of July 1980 a female Aedes sollicitans (Walker), was caught in the 
Blair Hills light trap located just south of Mishawaka in the central portion of the 



Entomology 277 

Table 2. Summary of New Jersey light traps collections, 1976-1980 







Year of collection 




















Five year 


Species 


1976 


1977 


1978 


1979 


1980 


total 


Aedes 

abserratus 


7 








1 


8 


aurifer 


- 


- 


2 


3 


31 


36 


canadensis 


12 


49 


509 


623 


28 


1221 


cinereus 


4 


24 


16 


194 


80 


318 


dorsalis 


6 


1 


11 


6 


74 


98 


excrucians 


25 


3 


5 


- 


11 


44 


fitchii 


- 


1 


9 


1 


22 


33 


flavescens 


- 


- 


- 


1 


- 


1 


sollicitans 


- 


- 


- 


- 


7 


7 


sticticus 


1592 


113 


818 


451 


51 


3025 


stimulans 


271 


237 


468 


6705 


2349 


10030 


triseriatus/hendersoni 


55 


141 


239 


345 


546 


1326 


trivittatus 


23 


180 


760 


328 


2362 


3653 


vexans 


9380 


8913 


40748 


42122 


138356 


239519 


Anopheles 














barberi 


3 


1 


4 


2 


3 


13 


crucians 


5 


- 


19 


- 


- 


24 


punctipennis 


469 


518 


897 


1840 


2216 


5940 


quadrimaculatus 


330 


376 


688 


795 


2067 


4256 


walkeri 


108 


62 


65 


492 


458 


1185 


Coquillettidia 














perturbans 


284 


699 


277 


1027 


2277 


4564 


Culex 














erraticus 


4 


1 


12 


48 


21 


86 


pipiens/restuans 


5238 


3262 


7088 


18141 


16424 


50153 


salinarius 


1005 


572 


135 


1058 


55 


2825 


tarsalis 


7 


88 


38 


16 


7 


156 


territans 


306 


305 


486 


1505 


1482 


4084 


Culiseta 














inornata 


10 


103 


336 


265 


85 


799 


melanura 


3 


1 


5 


2 


4 


15 


morsitans 


2 


4 


7 


2 


1 


16 


minnesotae 


- 


- 


5 


5 


8 


18 


Orthopodomyia 














alba/signifera 


24 


7 


8 


16 


34 


89 


Psorophora 














ciliata 


30 


20 


17 


43 


62 


172 


columbiae 


2 


8 


- 


114 


321 


445 


ferox 


2 


- 


1 


- 


1 


4 


Uranotaenia 














sapphirina 


203 


96 


897 


1141 


1570 


3907 


Undetermined 


16 


- 


2242 


413 


119 


2790 


Total 


19426 


15785 


56812 


77704 


171133 


340860 



county. Subsequently 6 additional specimens were collected from 5 different light 
traps representing almost the entire county (Figure 2). This species is a very 
strong flier, reportedly being able to disperse over 160 km (1), making it difficult to 
locate the larval sites. The large number of widespread collections indicate that 
this species is well established in the county. Ae. sollicitans is primarily a coastal 
mosquito breeding in water with a high salt content. It has, however, been 
reported from several sites as far inland as North Dakota and Arizona (3), usually 



278 



Indiana Academy of Science 




Figure 3. Counties from which Ae. flavescens, Ae. sollicitans and Ps. cyanescens 
have been recorded in Indiana. 



associated with some form of salt pollution. Ae. sollicitans had only been reported 
from two counties in Indiana previously, Posey and Vanderburgh, both in the ex- 
treme southwest corner of the state. Both of these sites were associated with high 
salinity. Recently Ae. sollicitans has been recorded from 6 additional counties in 
southern Indiana (Figure 3) and is probably present in most of the counties south of 
Indianapolis (M. Sinsko 1980, personal communication). In Ohio A e. sollicitans has 
been recorded in 19 counties (Figures 4) (R. Berry 1980, personal communication). 
Some of the larval sites in Ohio are run-off from fly ash dumps, salt mining opera- 
tions, brine pits near oil wells and industrial waste with high salinity. Ae. 



Entomology 



279 



sollicitans has become a locally serious pest near some of these sites. In addition 
this species is also a vector of Eastern Equine Encephalitis in eastern coastal 
endemic areas. The proximity of St. Joseph County to a known endemic focus of 
EEE in northwestern Indiana and southwestern Michigan is of concern. 

The ability of Ae. sollicitans to disperse over great distances and the increas- 
ing number of sites polluted by salt have allowed this species to expand its range 
from the coastal United States to the point where it can now be found in nearly 
every state east of the Rocky Mountains. 

Acknowledgement 

Funding for this study was provided by the St. Joseph County Health Depart- 
ment and NIH Research Grant No. AI-02753. 



Literature Cited 

1 . Carpenter, S. J. and W. J. LaCasse. 1955. Mosquitoes of North America. 
University of California Press, Berkeley, CA. 360 p. 




Figure 4. Distribution of Ae. sollicitans in Ohio. 



280 Indiana Academy of Science 

2 . Craig, G. B. Aedes atropalpus introduced into Indiana: Is this mosquito a 

potential vector of arboviruses? Proc. Ind. Vect. Cont. Assn. in press. 

3 . Knight, K. L. 1967. Distribution of Aedes sollicitans (Walker) and Aedes 

taeniorhynchus (Wiedemann) within the United States (Diptera: Culicidae). 
Georgia Ent. Soc. J. 2(1): 9-12. 

4 . Siverly, R. E. 1972. Mosquitoes of Indiana. Indiana State Board of Health, In- 

dianapolis, Indiana 126 p. 
5 . Shroyer, D. A., R. F. Beach, L. Munstermann, J. Peloquin, J. L. Petersen, 

R. P. Smith and D. B. Taylor. 1977. Mosquito diversity in St. Joseph County, 

Indiana (Diptera: Culicidae). Proc. Ind. Acad, of Sci. 86: 238-241. 
6 . Restifo, R. A. and G. C. Lanzaro. 1980. The occurrence of Aedes atropalpus 

(Coquillett) breeding in tires in Ohio and Indiana. Mosquito News 40(2): 

292-294. 



ENVIRONMENTAL QUALITY 

Chairman: Howard Dunn 

Department of Chemistry, 

Indiana State University at Evansville, 

Evansville, Indiana 47712 

Chairman-Elect: David Peterson 

Indiana State Board of Health, 

1330 West Michigan Street, 

Indianapolis, Indiana 46206 

ABSTRACTS 

Acid Precipitation Control Strategies. Thad J. Godish, Department of Natural 

Resources, Ball State University, Muncie, Indiana 47306. This investigation 

was designed to develop and to evaluate technically and politically various alter- 
native acid precipitation control strategies which primarily focus on SO x emissions 
to the atmosphere. SO x producing states of the Ohio River Basin and upper middle 
west were included in the model study area. SO x control strategies based on 
universal coal cleaning requirements, emission limitations, total atmospheric 
burden, and best available control technology are described and evaluated. 

Sensitivity of Weed Species to Ozone Exposures. Thad J. Godish, Department of 

Natural Resources, Ball State University, Muncie, Indiana 47306. Nine weed 

species common to the Midwest were exposed to concentrations of 0.15 and 0.25 
ppm ozone for 3 hours in order to determine their sensitivity. Based on plant 
response to their exposures, the test species were classified as sensitive, slightly 
sensitive, intermediate-resistant and resistant. Smartweed [Polygonum persicaria) 
and cocklebur (Xanthium ckinense) were rated as slightly sensitive with some 
plants showing symptoms at 0.15 ppm. Lambsquarters (Ckenopodium album) was 
intermediate in sensitivity. Pigweed (Amaranthus retroflexus) was intermediate- 
resistant showing slight injury after an expousre to 0.25 ppm. All other species 
tested including giant ragweed [Ambrosia trifida), dwarf ragweed (Ambrosia 
artemisiifolia), jimsonweed (Datura stramonium), intermediate dogbane 
(Apocynum medium), flower of an hour (Hibiscis trionium) and velvetleaf (Abutilon 
Theophrasti) showed no symptoms after exposure to 0.25 ppm ozone and were 
classified as resistant. 

Formaldehyde Air Pollution in Residential Housing. Thad J. Godish, Department 
of Natural Resources, Ball State University, Muncie, Indiana 47306, and Virgil 
Konopinski and David Peterson, Division of Industrial Hygiene, Indiana State 

Board of Health, Indianapolis, Indiana 46206. This report summarizes results 

of formaldehyde sampling in home interiors in Indiana conducted by the Indiana 
State Board of Health and Ball State University. In 26 cases investigated for- 
maldehyde levels ranged from 0.03 to 1.35 ppm. Sources of free formaldehyde in- 
cluded urea-formaldehyde foam insulation, particleboard underlayment, cabinets, 
paneling, and furniture. A range of health symptoms were reported by 
homeowners to be associated with their homes including eye and upper respiratory 
irritation, rhinitis, coughing, difficulty in breathing, sinusitis, headaches, dizziness, 
tiredness, difficulty in sleeping, digestive problems, chest and abdominal pains, and 
abnormal stool or diarrhea. 

281 



282 Indiana Academy of Science 

The Chemical Properties of Biological Sludges. Robert H. L. Howe, West 

Lafayette, Indiana 47906. The Chemical Properties of Anaerobic and Aerobic 

Biological Sludges are discussed. Their values and useful potentials are presented. 

The Meaning of pH in Environmental Control and Wastes Treatment. Robert H. 
L. Howe, West Lafayette, Indiana 47906. The meaning of pH and the computa- 
tion of the "average" pH values by the proper hydrogen ion concentration and the 
final pH of a number of acid and basic solutions are explained. The logarithmic ex- 
pression of the proper pH computation is emphasized. 

A Groundwater Quality Study of the Franklin County Sanitary Landfill, 

Brookville, Indiana. Patrick Sullivan, Department of Natural Resources, Ball 
State University, Muncie, Indiana 47306. The Franklin County Sanitary Land- 
fill is located in sand and gravel deposits within a meander of the Whitewater 
River. Given the high permeability of the geologic materials, this landfill site is not 
suitable for environmentally sound waste disposal. However, this circumstantial 
evidence does not prove that groundwater contamination is occurring. This uncer- 
tainty resulted in this project with the objective of identifying possible ground- 
water and/or surface water contamination. 

Monitoring wells (PVC Pipe) were installed at four sites about the landfill 
boundary. Over a period of five months water samples were retrieved from the 
monitoring wells as well as upstream and downstream of the landfill. These 
samples were analyzed for: pH, TDS, COD, CI, Cd, Zn, Pb, Ca, Ti, and Fe. 

The results of the analysis indicated the following: a) leachate is leaving the 
landfill and is being detected in the monitoring wells, b) only Fe exceeded EPA 
standards, and 3) the reason higher levels of all parameters were not recorded was 
probably due to dilution. 

The Effects of Acid Mists on Nitrogen-Fixing Blue-Green Algae. RONALD W. 
Usher and Edmund A. Schofield, The Institute of Ecology, Indianapolis, Indiana 

46208. Nitrogen-fixing blue-green algae are important components of the 

pioneer ecosystem on the foredunes of the Indiana Dunes State Park. They con- 
tribute combined nitrogen and reduced carbon to the soil, increase the soil's water- 
holding capacity, and provide a substratum suitable for the germination of seeds. 
Since blue-green algae require neutral to slightly alkaline conditions, it is signifi- 
cant that the pH of the precipitation currently falling on the dunes is 4.0 to 4.5. 
Because sand has a low buffering capacity, I hypothesized that acid precipitation 
may be inhibiting the growth of and nitrogen fixation by the blue-green algae grow- 
ing on the Indiana Dunes. To test my hypothesis, I studied the distribution and 
ecological relationships of blue-green algae in the Indiana Dunes State Park. 
Crusts of nitrogen-fixing blue-green algae grew abundantly on the foredunes. They 
consisted of eight species of blue-green algae. The pH of the foredune sand was 8.0, 
that of Lake Michigan water, 7.9. In the laboratory, I subjected unialgal cultures of 
Nostoc commune (UTEX B1621), and Scytonema hofmannii (UTEX B1834) (two of 
the species identified in the crusts), as well as samples of dune crusts, to mists of 
pH 3.5, 4.5, 5.5, 6.5, 7.5, 8.5, and 9.5 over a period of several weeks. The mists in- 
hibited nitrogen fixation by N. commune by as much as 90 percent, whereas they 
stimulated fixation by S. hofmannii. All of the N. commune cultures recovered 
from the effects of the mists, however. I obtained inconclusive results from the 
crusts collected at the Indiana Dunes, although they appeared to be stimulated by 
the treatments. 



Effects of Nitric Oxide, Nitrogen Dioxide and Nitric Oxide-Nitrogen Dioxide 
Pretreatments on the Sensitivity of Tomato cv. Rutgers to Ozone 

Thad J. Godish 

Department of Natural Resources, 

Ball State University, 

Muncie Indiana 47306 

Introduction 

Plant research with gaseous mixtures has revealed pollutant-plant inter- 
actions which profoundly effect plant response. These pollutant-plant interactions 
can be characterized as being synergistic (response significantly greater than ad- 
ditive effects of plants exposed to single pollutants), additive, or antagonistic 
(response less than additive effect of plants exposed to single pollutants). 

Studies of these interactions are of considerable importance since plants 
growing in the ambient environment (unlike those of most laboratory experiments) 
usually are exposed to more than one pollutant simultaneously or sequentially over 
a period of 24 hours. Laboratory research has indeed confirmed that simultaneous 
exposures to gaseous mixtures such as 3 and S0 2 can significantly effect plant 
response. (2, 4,-9). 

Most studies of interaction effects have focused on simultaneous exposures to 
gaseous mixtures. In many ambient atmospheric situations, however, concentra- 
tions of specific pollutants show fluctuations which are periodic, reaching max- 
imum levels at predictable hours of the day. In urban areas which have significant 
auto emissions, peak ozone levels occur at midday. This ozone peak is preceded by 
peaks in nitrogen dioxide (circa^. 10:00 am) and nitric oxide (circa 8:00 am). 
Therefore, under ambient conditions, plants may not only be exposed to more than 
one pollutant simultaneously, but are also exposed to some pollutants sequentially. 

This study was designed to determine the response of test plants to sequential 
exposures to nitric oxide, nitrogen dioxide and ozone at several exposure levels 
and sequences. Specific emphasis was placed on the determination of the effects of 
preexposure to nitric oxide, nitrogen dioxide and nitric oxide following by 
nitrogen dioxide on symptom development in plants subsequently exposed to 
phytotoxic levels of ozone. 

Materials and Methods 

Tomato seedlings, Lycopersicon esculentum cv. Rutgers were grown in 10 cm 
diameter plastic pots containing a 1:1:1 mixture of peat, perlite and loam soil sup- 
plemented with a 12:12:12 water soluble fertilizer. These seedlings were grown in 
an environmental chamber with a 27/17°C day/night temperature regime and a 
14-hour photoperiod. Relative humidity was uncontrolled ranging from 40-60% 
during the light period and 60-80% during the dark period. Cool white fluorescent 
and incandescent lamps provided a light intensity of 8.93 x 10 4 ergs cm' 2 sec * at the 
plant surface. Plants were refertilized with a 12:12:12 slow release granular fer- 
tilizer mix (Osmocote) 2 weeks after transplanting. Plants grown to the second 
nine-foliate state were utilized for all exposures. 

Exposures were conducted in two 10 cubic foot volume plexiglass exposure 
chambers. Air flow through the chambers was approximately 15 cubic feet per 

283 



284 Indiana Academy of Science 

minute. During exposures, exposure chamber temperature and relative humidity 
were maintained at 27 ± 2°Cand70 ± 2%, respectively. Light intensity of 10.7 x 
10 ~ 4 ergs ^ sec" * was provided by cool white fluorescent and incandescent lamps 
above the exposure chambers. Ozone was produced for all exposures by an Alron 
high voltage ozone generator. Ozone concentrations were monitored during 
exposure by a Mast oxidant meter calibrated with the Potassium Iodide-Boric Acid 
Method (3). Prepared nitric oxide (0.1%)-nitrogen and nitrogen dioxide 
(0.2°/o)-nitrogen gas mixtures were used for nitric oxide and nitrogen dioxide 
exposures, respectively. The desired exposure concentrations were achieved by fur- 
ther diluting these gas mixtures with room air which was drawn into the exposure 
chambers. Nitrogen dioxide concentrations were monitored by drawing a 30 minute 
six liter air sample through a bubbler containing absorbing reagent. The sample was 
analyzed by using the Sodium Arsenite Method (1). Nitric oxide concentrations were 
also monitored by the same method. For nitric oxidide determinations it was 
necessary to convert the nitric oxide into nitrogen dioxide by drawing the air sam- 
ple through a U-tube containing a chromium trioxide medium. 

As a part of the experimental design all plants used in this study were sub- 
jected to a phytotoxic exposure of 0.20 ppm/3 hours ozone. Half of these plants were, 
in addition, pretreated with non-phytotoxic exposures of nitric oxide and nitrogen 
dioxide at several exposure levels and sequences. These included (1) 1 ppm NO/3 hrs. 
(2) 0.35 ppm NO/3 hrs. (3) 1 ppm N0 2 /3hrs. (4) 0.35 ppm N0 2 /3 hrs and (5) 0.35 ppm 
NO/2 hrs. followed by 0.35 ppm N0 2 /2 hrs. Plants which were exposed to ozone but 
were not pretreated with nitric oxide or nitrogen dioxide served as controls. In 
preliminary exposures to the above levels of nitric oxide and nitrogen dioxide these 
gases were observed not to by phytotoxic, that is, no visible symptoms were produc- 
ed from such exposures. 

Because of limited chamber space available, only four tomato plants were 
exposed per chamber for a given exposure. One chamber was used for sequential 
exposures to nitric oxide, nitrogen dioxide and ozone; plants in the second chamber 
were only exposed to ozone. To provide sufficient numbers of plants for statistical 
evaluation, each of the five experiments was conducted three times and the data 
pooled. In each case sequential exposures were paired with exposures to ozone 
alone. 

After exposure, plants were returned to the environmental chamber. Symp- 
tom development was evaluated 96 hours after exposure. Total symptom severity 
was determined by the °/o leaf area exhibiting flecking, mottling and bifacial 
necrosis. In addition, symptom severity based on bifacial necrosis was also deter- 
mined as % of the leaf area injured. The data were statistically evaluated by 
employing Student's t-test. An alpha level of .05 was accepted as significant. 

Results 

The effects of nitric oxide, nitrogen dioxide, and nitric oxide followed by 
nitrogen dioxide pretreatments on symptom development in tomato plants which 
were subsequently exposed to a phytotoxic ozone dose are summarized in Table 1. 
Plants pre-exposed to nitric oxide, 0.35 ppm/3 hrs. (experiment 2) and nitrogen diox- 
ide, 1.00 ppm/3 hrs. (experiment 3) were observed to show a significant antagonistic 
effect on symptom development in plants which were later exposed to phytotoxic 
ozone levels. On the other hand, no significant differences were observed in ex- 
periments 1 (1.00 ppm NO/3 hrs.), 4 (0.35 ppm N0 2 /3hrs.) and 5 (0.35 ppm NO/2 hrs. 
followed by 0.35 ppm N0 2 /2 hrs.). The apparent antagonistic effects as indicated in 



Environmental Quality 



285 



Table I. Effect of nitric oxide, nitrogen dioxide, and nitric oxide - nitrogen 
dioxide pretreatments on ozone-induced foliar injury on tomato. N = 12 



Experi- 






Pollutant concentration (ppm) 




% leaf area 


ment 






and 


exposure 


duration 




injured 




NO 


hrs. 


N0 2 


hrs. 


°3 


hrs. 




1 


1.00 


3 


_ 


_ 


0.20 


3 


15.4 




- 


- 


- 


- 


0.20 


3 


24.0 


2** 


0.35 


3 


_ 


_ 


0.20 


3 


27.9 




- 


- 


- 


- 


0.20 


3 


37.7 


3* 


_ 


_ 


1.00 


3 


0.20 


3 


25.7 




- 


- 


- 


- 


0.20 


3 


45.0 


4 


_ 


_ 


0.35 


3 


0.20 


3 


16.5 




- 


- 


- 


- 


0.20 


3 


17.9 


5 


0.35 


2 


0.35 


2 


0.20 


3 


10.9 




- 


- 


- 


- 


0.20 


3 


18.0 



the means of experiments 1 and 5 were not statistically significant at the 0.05 alpha 
level. 

Results presented in Table I are based on total leaf area injured which included 
symptoms of flecking, mottling, and bifacial necrosis. When the effect on the 
development of bifacial necrosis, the severest form of injury, was considered results 
similar to those reported in Table I were obtained. 



Discussion 

Results of experiments 2(0.35 ppm NO/3 hrs.) and 3 (1.00 ppm N0 2 /3 hrs.) in- 
dicate that pretreatment with gases such as nitric oxide and nitrogen dioxide 
which themselves do not induce visible injury, can antagonize the phytotoxic ef- 
fects of ozone in sequential exposures. These results however, were not supported 
by the results of experiments 1 (1.00 ppm NO/3 hrs.), 4 (0.35 ppm N0 2 /3 hrs.) and 5 
(0.35 ppm NO/2 hrs followed by 0.35 ppm N0 2 /2 hrs.). The results of experiment one 
are particularly anomalous, as this exposure did not result in a statistically signifi- 
cant antagonistic response whereas exposure to a smaller dose, 0.35 ppm NO/3 hrs. 
did. On the other hand, the higher dose of N0 2 , experiment 3 resulted in an an- 
tagonistic response and the smaller dose did not. Based on dose-response con- 
siderations the results of experiments 3 and 4 could be expected but those of 1 and 
2 are anomalous. 

What is the basis of the anomaly observed in NO pretreatments? It is prob- 
able that this is due at least in part to the large variability in data collected. The 
large data variability in these experiments may have been due to the limitations 
placed on the study by exposure chamber capacity. To provide sufficient numbers 
for statistical evaluation 3 exposures were conducted for each experiment. These 
exposures were conducted on different days. The variation in symptom response 
from day to day as evidenced in control plants was considerable despite attempts 
to minimize this day to day variation by controlling preexposure environmental 
growth and exposure conditions. 

Antagonistic effects of pollutant gases have been previously reported by 
Heagle and Johnston (2). In studies with soybean Heagle and Johnston observed 
that simultaneous exposure to mixtures of S0 2 and 3 could result in an an- 



286 Indiana Academy of Science 

tagonistic effect when injury from either gas singly was severe. They suggested 
that the 3 in the exposure mixtures of S0 2 + 3 could at times protect soybeans 
from S0 2 . The observation by Heagle and Johnston that antagonism was observed 
only when plant injury was severe was evaluated in this study by assessing leaf 
area which exhibited the severest symptoms, bifacial necrosis. Similar results 
were obtained when symptom development was evaluated on °/o leaf area injured 
by bifacial necrosis and total °/o leaf area injured. Apparently then antagonism in 
the study reported here was not related to symptom severity. 

Unlike the studies of Heagle and Johnston which utilized simultaneous ex- 
posures to phytotoxic gases, this study utilized sequential exposures to non- 
phytotoxic levels of NO and N0 2 followed by an exposure to phytotoxic levels of 
ozone. In experiments 2 and 3 such exposures apparently produced an antagonistic 
effect on ozone-induced symptom developed. What is the nature of this an- 
tagonism? Antagonistic mechanisms could include physiological changes in NO and 
N0 2 exposed plants such as partial stomatal closure and/or biochemical changes 
which protect plants from ozone. No experimental evidence on antagonistic 
mechanisms is, however, available. 

Literature Cited 

1. Christie, A. A., R. E. Lidzey, and D. W. F. Radford. 1970. Field Methods for 
the Determination of Nitrogen Dioxide in Air. Analyst 95: 519-524. 

2. Costonis, A. C. 1970. Acute Foliar Injury of Eastern White Pine Induced by 
Sulfur Dioxide and Ozone. Phytopathology 60: 994-999. 

3. Flamm, D. L. 1977. Analysis of Ozone at Low Concentrations with Boric Acid 
Buffered KI. Environ. Sci. Technol. 11:978-983. 

4. Heagle, A. S. & J. W. Johnston. 1979. Variable Responses of Soybeans to 
Mixtures of Ozone and Sulfur Dioxide. JAPCA 29(7): 729-732. 

5. Jacobson, J. S. and L. J. Cola veto. 1976. The combined Effect of Sulfur Diox- 
ide and Ozone on Bean and Tobacco Plants. Environ. Ex. Bot. 16: 277. 

6. MacDowell, F. D. H., and A. F. W. Cole. 1971. Threshold and Synergistic 
Damage to Tobacco by Ozone and Sulfur Dioxide. Atmos. Environ. 5: 553-559. 

7. Menser, H. A. and H. E. Heggestead. 1966. Ozone and Sulfur Dioxide 
Synergism: Injury to Tobacco Plants. Science 153: 424-425. 

8. Tingey, D. T., R. A. Reinert, J. A. Dunning and W. W. Heck. 1973. Foliar In- 
jury Responses of Eleven Plant Species to Ozone/Sulfur Dioxide Mixtures. At- 
mos. Environ. 7:201. 

9. Tingey, D. T. and R. A. Reinert, 1975. The Effect of Ozone and Sulfur Dioxide 
Singly and in Combination on Plant Growth. Environ. Pollut. 9: 117. 



Phosphorus Status of Selected Indiana Lake and Reservoir Sediments 

D. W. Nelson and E. D. Orme 

Department of Agronomy, 

Purdue University, 

West Lafayette, Indiana 47907 

Introduction 

Eutrophication is the natural aging process of lakes that is characterized by 
increasing organic productivity and accelerated by excessive nutrient input. There 
is good evidence to show that phosphorus (P) is a limiting nutrient for algal growth 
in many lake systems and the key element in the eutrophication process 
(Vollenweider, 1971; Syers et al., 1973). Soluble inorganic phosphorus (SIP) is the 
major form of P taken up by aquatic plants and, thus, the P form most responsible 
for eutrophication. This form of P is also involved in equilibrium sorption- 
desorption reactions with lake sediments. Sediments may serve as sinks for SIP 
present in lake water or sources of SIP for algal uptake depending upon the nature 
of the equilibrium. It is apparent from studies on Wisconsin lakes that the adsorp- 
tive site for SIP is an aluminum and iron rich amorphous gel complex. This appears 
to be true for both calcareous and noncalcareous sediments. (Williams et al., 1971a; 
1971b). 

The sorption-desorption equilibrium of P, as affected by sediment and lim- 
nological conditions, determines the ultimate concentration of SIP maintained in 
the water phase by sediment. The concentration of SIP in the water phase at which 
P is neither sorbed or desorbed from the sediment phase is a unique property of a 
sediment known as equilibrium phosphate concentration (EPC). It has been shown 
that when sediment and phosphate solutions were equilibrated in the laboratory 
there was desorption of sediment P at low concentrations of added P, whereas at 
higher SIP concentrations P was sorbed by the sediment. The EPC value is the 
point of zero sorption and is determined from a plot of SIP concentration at 
equilibrium versus P sorbed. 

Little is known about the P status of Indiana lake and reservoir sediments. 
Therefore, the objectives of this study were: to survey the forms of P in selected 
lake and reservoir sediments, to determine the nature of P exchange between sedi- 
ment and water phases, and to relate sediment characteristics to existing water 
quality. 

Materials and Methods 

Wet sediment samples were collected by an Ekman dredge from the deepest 
portion of lakes and reservoirs, passed through a 4-mesh sieve, and thoroughly 
mixed. Sediments were stored as slurries in sealed glass containers at 4°C until 
analyzed. Solids content of sediments was determined by drying at 105°C and pH 
was estimated by glass electrode. An amount of wet sediment equivalent to 0.5 g 
oven-dry weight was used in all sorption and extraction studies. Sorption and 
release of P by sediments was studied by shaking each sediment with 40 ml of 
0.01M calcium chloride solution containing from to 400 fig P/ml as monopotassium 
phosphate for 24 hours, centrifuging the mixture for 10 minutes at 12,000 x g, filter- 
ing the supernatant through Whatman 2V paper, and determining SIP in the 
filtrate by the method of Murphy and Riley (1962). The amount of SIP sorbed or 

287 



288 Indiana Academy of Science 

desorbed was calculated as the difference between the amount added and the 
amount remaining in solution at equilibrium. Amounts of SIP sorbed by the sedi- 
ment were plotted against the equilibrium concentrations of SIP to permit 
graphical estimation of EPC for each sediment (Taylor and Kunishi, 1971). Data ob- 
tained from sorption studies were also plotted in the form of Langmuir adsorption 
isotherms from which adsorption maxima, buffering capacity, and binding energy 
values were calculated (Syers et al., 1973). 

Availability of P in sediments was estimated by extraction with reagents 
known to selectively remove P associated with specific components. Aluminum- 
bound P was determined by the 0.5M ammonium fluoride (pH 8.2) extraction 
method described by Jackson (1970). Phosphorus associated with amorphous iron 
oxides was estimated by the extraction with ammonium oxalate-oxalic acide 
reagent (Saunders, 1965) and P bound to amorphorus plus crystalline iron oxides 
was determined by the 0.5M sodium hydroxide extraction method described by 
Jackson (1970). Calcium-bound P was determined by extraction of sodium 
hydroxide-treated samples with 0.5N sulfuric acid as described by Jackson (1970). 
The amounts of labile P present in selected sediments was estimated by the 32 P 
isotopic dilution technique (Olson and Dean, 1965). 

Subsamples of wet sediment were air-dried, passed through a 2 mm sieve and 
subjected to particle size analysis (Kilmer and Alexander, 1946). Air-dry sediment 
was further ground to < 80-mesh and analyzed for total N (Nelson and Sommers, 
1972), total P (Sommers and Nelson, 1972), organic carbon (Mebius, 1960), and in- 
organic carbon (Allison, 1960). Data reported are on an oven-dry basis and repre- 
sent the average of samples collected from a particular lake or reservoir. Methods 
described by Steel and Torrie (1960) were used for correlation analyses. 

Results and Discussion 

Table 1 provides data on the general characteristics of the lake and reservoir 
sediments studied. There were large differences in composition between lake and 
reservoir sediments likely because of variations in watershed conditions, sediment 
parent material, and age of the impoundments. Lake sediments contained much 
higher levels of organic carbon (ave. of 12.3% versus 2.1%), inorganic carbon (ave. 
of 5.9% versus 0.4%), and total nitrogen (ave. of 0.83% versus 0.18%) than did 
reservoir sediments. Reservoir sediments contained higher concentrations of clay- 
size material than did lake sediments (an ave. of 28 and 7%, respectively). 

Table 1. General characteristics of lake and reservoir sediments. 







Organic 


Inorg. 


Total 






Sediment 


Code 


C 


C 


N 


Clay 


pH 














Lakes: 














Big Turkey 


BT 


8.2 


7.8 


0.53 


12 


7.8 


Cedar 


Cd 


13.7 


1.1 


0.90 


5 


7.6 


Maxinkuckee 


Mx 


9.0 


8.5 


0.64 


8 


7.8 


Palestine 


Pa 


10.1 


6.7 


0.86 


7 


7.7 


Sylvan 


Sy 


13.0 


5.1 


0.92 


7 


7.5 


Wawasee 


Wa 


19.6 


6.2 


1.13 


5 


7.6 


Reservoirs: 














Cataract 


Ca 


1.9 


0.6 


0.16 


16 


7.8 


Mansfield 


Mn 


1.9 


0.5 


0.14 


40 


7.5 


Mississinewa 


Mi 


2.1 


0.4 


0.19 


38 


8.1 


Monroe 


Mo 


2.3 


0.0 


0.21 


17 


7.6 



Environmental Quality 289 

Table 2. Phosphorus components present in lake and reservoir sediments. 





Total 


Organic 




Inorganic P 


extracted by 














Sediment 


P 


P 


NH 4 F 


Oxalate 


NaOH 


H 2 S0 4 + 




Mg/g 
690 












BT 


36 


7 


23 


5 


58 


Cd 


770 


51 


16 


15 


12 


35 


Mx 


470 


52 


11 


14 


4 


43 


Pa 


1150 


56 


8 


10 


7 


36 


Sy 


1290 


42 


8 


16 


9 


49 


Wa 


570 


53 


10 


14 


4 


41 


Ca 


1170 


44 


13 


24 


22 


29 


Mn 


1080 


48 


20 


22 


18 


29 


Mi 


740 


47 


27 


23 


21 


29 


Mo 


630 


48 


10 


19 


30 


19 



+ Extraction with H z S0 4 after NaOH extraction was completed. 

However, pH values of lake and reservoir sediments were similar. Reservoir 
sediments tended to have compositions similar to the soils in the watershed, 
whereas lake sediments were very different from the soils surrounding the lake. 
All lake sediments contained significant amounts of free carbonates, but only small 
amounts of carbonates were found in reservoir sediments. 

Table 2 provides data on the P components present in sediments. Lake and 
reservoir sediments contained similar levels of total P (an ave. of 823 and 905 /xglg, 
respectively) and organic P (an ave. of 395 and 425 figlg, respectively. A higher pro- 
portion of total P was present as aluminum-bound and amorphorus iron-bound P in 
reservoir sediments as compared to lake sediments. This finding is important 
because these P components are thought to be the most available for release to the 
water phase. Furthermore, the amount of sodium hydroxide-extractable P has 
been used as an index of the algal available P content of a sediment. Oxalate and 
sodium hydroxide extracted similar amounts of P from reservoir sediments sug- 
gesting that limited amounts of P are associated with crystalline iron oxides in 
these sediments. Surprisingly, oxalate extracted significantly more P from most 
lake sediments than did sodium hydroxide. The proportions of total P present as 
calcium-bound P (sulfuric acid extractable) were much higher for lake sediments 
(average of 44%) as compared to reservoir sediments (average of 27%). Calcium- 
bound P is thought to be largely unavailable for release to the water column. 

Eutrophication indices for each lake and reservoir studied are given in Table 
3. These indices were supplied by the U.S. Environmental Protection Agency and 
are based on extensive examination of water quality and productivity in the im- 
poundments. In general, the lakes tended to have higher indices than the reser- 
voirs indicating more advanced eutrophication in the former. Cedar, Sylvan, and 
Palestine Lakes were judged to be eutrophic, whereas Lake Maxinkuckee was 
probably oligotropic. All reservoirs studied had relatively low eutrophication in- 
dices in the mesotrophic range. 

Table 3 also gives data on the EPC, P adsorption maxima, and labile P values 
for sediments. The EPC values for reservoir sediments were low (average of 0.003 
Hg P/ml), whereas the lake sediments had relatively high EPC levels (average of 
0.032 ng P/ml). This suggests that lake sediments had the capacity to buffer the SIP 
concentration in the water phase more than ten times higher than reservoir 
sediments. Big Turkey, Cedar, and Sylvan lake sediments had particularly high 
EPC values. On the average, reservoir sediments had higher capacities for adsorp 



290 



Indiana Academy of Science 



Table 3. Eutrophication indices of impoundments and EPC, phosphorus 
adsorption maxima, and labile P values for sediments. 



Sediment 



Eut. 
Index 



EPC 



P 
lbs. max. 



Labile 
P 



BT 
Cd 
Mx 
Pa 

Sy 
Wa 
Ca 
Mn 
Mi 
Mo 





Aig/ml 


^g/g 


Mg/g 


42 


0.042 


3320 


154 


65 


0.067 


2420 


- 


18 


0.006 


2630 


- 


60 


0.020 


6940 


- 


62 


0.053 


1870 


- 


23 


0.004 


3700 


- 


26 


0.004 


7500 


- 


21 


0.001 


8350 


89 


20 


0.001 


3530 


- 


32 


0.004 


5020 


- 



tion of P than did lake sediments likely because of the higher levels of amorphous 
iron and aluminum in the former (Orme and Nelson, 1979). About 8 and 22% of the 
total P in Mansfield and Big Turkey sediments, respectively, was isotopically ex- 
changeable. However, 18 and 5% of the total P in Mansfield and Big Turkey 
sediments, respectively, was extractable with sodium hydroxide. These findings in- 
dicate that the two methods (i.e. sodium hydroxide extraction and 32 P exchange) 
used for estimating available P measure two different P pools in sediments. 

Table 4 presents data on the relationships between eutrophication indices of 
impoundments and sediment P component concentrations. There was a direct cor- 
relation between the eutrophication index of lakes and reservoirs and the EPC 
values of the sediments (Figure 1). This finding indicates that sediment EPC is an 
important water quality determinant and suggests a direct role of sediments in 
supplying SIP to the water column. Eutrophication indices of lakes were also 
directly related to the levels of total P, oxalate-extractable P, ammonium fluoride- 
soluble P, and NaOH-extractable P in sediments, but reservoir eutrophication in- 
dices were not correlated with levels of these P components in sediments. When all 
impoundments were considered, eutrophication indices were related to levels of 
sulfuric acid-soluble P in sediments, however, no such relationship was observed 
when only lakes or reservoirs were considered separately. 

Table 4. Relationships impoundment eutrophication indices and sediment P 

components and between sediment EPC values and 

other P components of sediments. 





Variable 2 






Sediments considered: 




Variable 1 


Lake 


Reservoir 


All 










r> tt 




Eut. index 


EPC 


+ 


.81* 


+ .89 


+ .87* 


Eut. index 


Total P 


+ 


.81 


NS 


NS 


Eut. index 


Oxalate P 


+ 


.67 


NS 


NS 


Eut. index 


NH 4 F-P 


+ 


.86 


- .98 


NS 


Eut. index 


NaOH-P 


+ 


.94* 


NS 


NS 


Eut. index 


H 2 S0 4 -P 




NS 


NS 


+ .62 


EPC 


Total P 




NS 


NS 


NS 


EPC 


Oxalate P 


+ 


.69 


NS 


NS 


EPC 


NH 4 F-P 


+ 


.73 


- .84 


NS 


EPC 


NaOH-P 


+ 


.75 


NS 


NS 


EPC 


H,S0 4 -P 




NS 


NS 


+ .55 



'Significant at 99% confidence level, NS, not significant at 90°/o confidence level. 



Environmental Quality 



291 



X 

a 



o 



75 



60 



y 45 



CE 

o 

X 

a. 
o 

C* 

h- 
3 



30 



15 







Figure 1. 



o LAKE R = 

x RESERVOIR 



81 

R = .89 







.02 
EPC, 



.04 
UG P/ML 



.06 



08 



Relationship between impoundment eutrophication indices and 
sediment EPC values. 



Table 4 also gives data on the relationships between EPC values and levels of 
sediment P components. Amounts of oxalate-, ammonium fluoride, and sodium 
hydroxide-extractable P in lake sediments were directly correlated with EPC 
values. No P component in reservoir sediments was directly related to EPC levels. 
When all sediments were considered the EPC values were positively correlated 
with the levels of sulfuric acide-soluble P. The significance of the latter finding is 
difficult to assess and it may result from the fact that lake sediments had higher 
EPC values and levels of sulfuric acid-extractable P without indicating a cause- 
effect relationship. 



Conclusions 

The following conclusions may be drawn from this work: (1) Lake and reser- 
voir sediment have very different levels of clay, organic carbon, inorganic carbon, 
and total nitrogen, and consequently, differ in P retention capacity, (2) Reservoir 
sediments have P components similar to soil, whereas lake sediments have reduced 



292 Indiana Academy of Science 

levels of iron and aluminum-bound P and increased levels of calcium-bound P, (3) 
From 7 to 23% of total sediment P is potentially available to algae, (4) Reservoir 
sediments appear to be sinks for P, but lake sediments may be P sources or sinks 
depending upon SIP concentration in the water column, (5) Eutrophication indices 
of lakes and reservoirs are directly related to sediment EPC values suggesting the 
involvement of sediment in biological productivity and overall water quality in im- 
poundments. 

Literature Cited 

1. Allison, L. E. 1960. Wet-combustion apparatus and procedure for organic and 
inorganic carbon in soil. Soil Sci. Soc. Am. Proc. 24:36-40. 

2. Jackson, M. L. 1970. Soil Chemical Analysis. Prentice-Hall, Inc. Englewood 
Cliffs, N.J. pp. 162-162.9. 

3. Kilmer, V. J. and L. T. Alexander. 1949. Methods of making mechanical 
analysis of soils. Soil Science 68:15-24. 

4. Mebius, L. J. 1960. A rapid method for determination of organic carbon in 
soils. Anal. Chim. Acta 22:120-124. 

5. Murphy, J. and J. P. Riley. 1962. A modified single solution method for the 
determination of phosphate in natural waters. Anal. Chim. Acta 27:31-36. 

6. Nelson, D. W. and L. E. Sommers. 1972. A simple digestion procedure for 
estimation of total nitrogen in soils and sediments. J. Environ. Quality 
1:423-425. 

7. Olson, S. R. and L. A. Dean. 1965. Phosphorus. In C. A. Black (ed.) Methods of 
Soil Analysis, Part 2. American Society of Agronomy, Madison, Wis. pp. 
1035-1049. 

8. Orme, E. D. and D. W. Nelson. 1979. Chemistry of phosphorus, cadmium, 
nickel, lead, and zinc in Indiana lake and reservoir sediments. Technical 
Report No. 122. Purdue Univ. Water Resources Center. 116 p. 

9. Saunders, W. M. H. 1965. Phosphate retention by New Zealand soils and its 
relationship to free sequioxides, organic matter, and other soil properties. 
New Zealand J. Agric. Res. 8:30-57. 

10. Sommers, L. E. and D. W. Nelson. 1972. Determination of total phosphorus in 
soils: a rapid perchloric acid digestion procedure. Soil Sci. Soc. Am. Proc. 
36:902-904. 

11. Steel, R. G. D. and J. H. Torrie. 1960. Principles and Procedures of Statistics. 
McGraw-Hill Book Co., Inc., New York. 481 p. 

12. Syers, J. K., R. F. Harris, and D. E. Armstrong. 1973. Phosphate chemistry in 
lake sediments. J. Environ. Quality 2:1-14. 

13. Taylor, A. W. and H. M. Kunishi. 1971. Phosphate equilibria on stream sedi- 
ment and soil in a watershed draining an agricultural region. J. Agr. Food 
Chem. 19:827-831. 

14. Vollenweider, R. A. 1971. Scientific fundamentals of the eutrophication of 
lakes and flowing waters, with particular reference to nitrogen and 
phosphorus as factors in eutrophication. Organization for Economic Coopera- 
tion and Development, Paris, France. 

15. Williams, J. D. H., J. K. Seyers, R. F. Harris, and D. E. Armstrong. 1971a. 
Fractionation of inorganic phosphate in calcareous lake sediments. Soil Sci. 
Soc. Am. Proc. 35:250-255. 

16. Williams, J. D. H., J. K. Seyers, D. E. Armstrong, and R. F. Harris. 1971b. 
Characterization of inorganic phosphate in noncalcareous lake sediments. Soil 
Sci. Am. Proc. 35:556-561. 



Chemical Analysis of Water Samples from Three Lakes in the Greene-Sullivan 
State Forest and the Wabash River at Clinton, Indiana 

Joseph R. Siefker and David P. McCleary 

Department of Chemistry 

Indiana State University 

Terre Haute, Indiana 47809 

Introduction 

Chemical analyses were performed on samples of surface waters of three 
typical lakes in the Greene-Sullivan State Forest to ascertain the quality of the 
water in those lakes, to determine concentrations of substances not listed in 
available publications, and to compare results of the determinations for the three 
lakes to that of the Wabash River. Further, the determinations for the Wabash 
River were compared to results obtained in earlier published findings to determine 
if the quality of water was deteriorating or improving. 

All three lakes are "strip-pit" lakes resulting from the natural filling of sur- 
face mining operations in the area. The common names of the lakes are Thousand 
Islands, Graveyard, and Airline Lake. 

Experimental 

Samples were taken by simply throwing a bucket into the lakes and retrieving 
the bucket as quickly as possible. Sites on the lakes were chosen so that natural 
mixing of the lakes insured a well-mixed sample. 

The Wabash River samples were taken at a point approximately 100 meters 
south of the bridge on S.R. 163 in Clinton, Indiana. 

Generally the methods of analysis used were from the publications of the 
American Public Health Association (1) and the Environmental Protection Agency 
(2). Chloride and fluoride ion selective electrodes were used with an Orion Model 
801 Analyzer to determine the activities of chloride and fluoride ions. Both deter- 
minations were performed within 24 hours of sample collection. Sodium and 
calcium ion concentrations were determined using a Jarrell-Ash Model 82-270 
Atomsorb atomic absorption spectrophotometer. All other substances were deter- 
mined by colorimetric methods. 

All samples were collected on Sunday and the determinations were performed 
on the following Monday except for sodium and calcium which were determined on 
two six-week intervals. 

Results and Discussion 

Samples were collected twelve times from September 7 through November 
23, 1975. Averages of the twelve samples are given in the following paragraphs. 

The concentration of aluminum seemed to have little or no correlation to rain- 
fall at any of the sites. The concentration in the Wabash River was the highest (0.08 
ppm), followed by Thousand Islands (0.07 ppm), Graveyard (0.03 ppm), and Airline 
(0.01 ppm). 

The concentration of ammonia was considerably greater in the three lakes 
than in the Wabash River. The greatest concentration was in Airline Lake (6.98 
ppm), followed by Graveyard (4.64 ppm), Thousand Islands (2.44 ppm), and the 

293 



294 Indiana Academy of Science 

Wabash River (0.69 ppm). The high concentrations in these lakes may indicate an 
absence of bacteria to oxidize the ammonia to nitrate. Another factor may be the 
reduction of nitrate to ammonia by metallic materials (perhaps iron, a common ele- 
ment in coal-containing soil). The results for the Wabash River (.69) are slightly 
higher than those given by Harmeson and Larson (3) in 1966. Their value for am- 
monia was 0.38 ppm. The value of ammonia was generally higher in all cases when 
rain was noted in the preceeding 72 hours. 

The concentrations of calcium were greatest in the three lakes. Airline Lake 
had the highest (116.8 ppm) followed by Graveyard (91.7 ppm), Thousand Islands 
(78.0 ppm), and the Wabash River (53.6 ppm). As noted, in general, by an ongoing 
survey of ground-water resources by the Indiana Department of Conservation 
Division of Water Resources, the waters of Greene (6) and Sullivan (7) counties tend 
to contain large amounts of calcium and magnesium caused by the composition of 
the underlying rock in those counties. The value of calcium reported by Harmeson 
and Larson (3) in 1966 for the Wabash River averaged 75 ppm. Little correlation 
between rainfall and calcium concentrations was seen. 

The chloride concentration for the Wabash River was highest at 18.7 ppm, 
followed by Thousand Islands (13.1 ppm), Graveyard (12.5 ppm), and Airline (11.2 
ppm). The level of chloride as reported by the Indiana State Board of Health in 1972 
(4) was 24 ppm at Terre Haute. Harmeson and Larson's value (3) in 1966 was 27 ppm 
for the Wabash River near Riverton, Indiana. Values for chloride listed in (6) and (7) 
for Greene and Sullivan counties are approximately 10-13 ppm, in excellent agree- 
ment with these results. In general, the overall results for chloride are in agree- 
ment with earlier work, within experimental error. 

The concentrations of copper was highest in the Wabash River (0.53 ppm), 
followed by Thousand Islands (0.27 ppm), Graveyard (0.12 ppm) and Airline (0.06 
ppm). 

The fluoride concentration of Airline Lake was the highest (0.391 ppm), followed 
by the Wabash River (0.304 ppm), Graveyard (0.265 ppm), and Thousand Islands 
(0.237 ppm). The value for fluoride reported by Lesniak, Tavenner and Siefker in 
1972 (5) was 0.25 ppm for the Wabash River at Terre Haute. The fluoride ion con- 
centration showed an inverse relationship to amounts of rainfall. 

The concentration of iron was highest in the Wabash River (1.16 ppm), followed 
by Airline (0.93 ppm), Thousand Islands (0.44 ppm), and Graveyard (0.37 ppm). The 
values reported in earlier data (3, 4, 5) are in good agreement with this value for the 
Wabash River. Also the values for iron reported in (6) and (7) for Greene and 
Sullivan counties are in good agreement with the values for the three lakes. The 
iron content of the samples was generally higher after rainfall. 

The nitrate concentration in the Wabash River was greatest (22.9 ppm) followed 
by Graveyard (15.2 ppm), Airline (14.0 ppm), and Thousand Islands (11.8 ppm). 
Again agreement between this value and earlier reports for the Wabash River (3, 
5) is excellent. One reason for the higher concentrations of nitrate in the Wabash 
River is probably from the use of nitrate fertilizers on the farmland drained by the 
Wabash River. 

Likewise, the concentration of nitrite was highest for the Wabash River (0.083 
ppm). The concentrations of nitrite in Thousand Islands (0.064 ppm), Graveyard 
(0.056 ppm), and Airline (0.051 ppm) were all lower. No correlation between rainfall 
and concentration of nitrite was discernable. 

The concentration of sodium was found to be higher in the Wabash River (54.6 



Environmental Quality 295 

ppm) than had previously been reported (3, 5). The earlier reports indicated values 
of about 20 ppm. The values for sodium concentrations in the lakes were lower than 
those for the Wabash River. The value for Thousand Islands (30.7 ppm) was 
highest, followed by Graveyard (28.4 ppm), and Airline (17.6 ppm). Little correla- 
tion between rainfall and concentration of sodium was noted. 

The phosphate concentration of the Wabash River (0.86 ppm) was highest, 
followed by Graveyard (0.73 ppm), Thousand Islands (0.68 ppm), and Airline (0.25 
ppm). The value of phosphate in the Wabash River in 1972 as reported by the In- 
diana State Board of Health (4) was 1.1 ppm. In general, the phosphate level was 
slightly higher after rainfall. 

The concentration of surfactants (methylene blue active substances) was 
highest in the Wabash River (0.23 ppm) followed by Thousand Islands (0.22 ppm), 
Graveyard (0.14 ppm), and Airline (0.14 ppm). 

The analyses showed a higher concentration of most substances in the 
Wabash River. The least amount of substances generally was found in Airline 
Lake. The overall results agree fairly well with earlier work with a few specific ex- 
ceptions. The known data indicate that the condition of the Wabash River is 
relatively stable, and that for the substances determined the concentrations are 
within limits set for drinking water. 

Literature Cited 

1. American Public Health Association. 1971. Standard methods for the ex- 
amination of water and wastewater, 13th edition, Washington, DC 874 p. 

2. Environmental Protection Agency. 1971. Methods of chemical analysis of 
water and wastes. Water Quality Office, Analytical Quality Control Lab., Cin- 
cinnati, OH 312 p. 

3. Harmeson. R. H., and T. E. Larson. 1969. Quality of surface water in Illinois, 
1956-1966. 111. State Water Sur. Bull. 54:175-180. 

4. Indiana State Board of Health and Stream Pollution Control Board. 
1972. Indiana water quality:monitor station records-rivers and streams. 116 p. 

5. Lesniak, D. G., M. C. Tavenner, and J. R. Siefker. 1973. Quantitative 
chemical analysis of specific components of the waters of Lost Creek and the 
Wabash River, Vigo County, Indiana. Proc. Indiana Acad. Sci. 82:176-179. 

6. Watkins, F. A., and D. G. Jordan. 1962. Ground-water resources of west- 
central Indiana, preliminary report, Greene County. Bulletin no. 11 of the In- 
diana Department of Conservation Division of Water Resources. 343 p. 

7. Watkins. F. A., and D. G. Jordan. 1962. Ground-water resources of west- 
central Indiana, preliminary report, Sullivan County. Bulletin no. 14 of the In- 
diana Department of Conservation Division of Water Resources. 255 p. 



GEOLOGY AND GEOGRAPHY 

Chairman: Kenneth R. Brehob 

Department of Earth Sciences, University of Notre Dame, 

Notre Dame, Indiana 46556 

Chairman-Elect: Henry H. Gray 

State Geological Survey, 611 North Walnut Grove, 

Bloomington, Indiana 47405 

ABSTRACTS 

The pH of Precipitation in Indianapolis. Konrad J. Banaszak, Office of Surface 
Mining, Indianapolis, Indiana 46204. The acid pH of precipitation in the north- 
eastern United States has been a matter of some concern. An automatic precipita- 
tion collector was located on the campus of IUPUI at 949 West New York Street, 
Indianapolis to ascertain the pH and anionic composition of precipitation. From 
September of 1978 to August of 1979, the pH of 73 individual precipitation events 
averaged 4.58 with a standard deviation of ± 0.59. The other measured consti- 
tuents averaged and had standard deviations of 4.16 ppm ± 1.51 for sulfate, 1.61 
ppm ± 1.63 for nitrate, and 0.89 ppm ± 0.79 for ammonium. No correlation of pH 
with either sulfate or nitrate was found. Nor was there any apparent correlation of 
the chemistry of the precipitation with the chemistry of the lower atmosphere as 
measured by the Environmental Protection Agency at the two sites within 0.1 
kilometer of the precipitation collecter. Although this study lasted only one year 
and lacks cationic data, it would appear that urban atmospheres, being more com- 
plex than rural atmospheres, have more complex patterns of acidic precipitation. 

A Social Impact Assessment of the Brookville Reservoir on Union County, In- 
diana. Thomas J. Gallagher, Margaret M. Kimmel, and Gary W. Barrett, In- 
stitute of Environmental Sciences, Miami University, Oxford, Ohio. The Brook- 
ville Reservoir, located in Franklin and Union Counties, Indiana, has already 
manifested both positive and negative impacts on the residents of this area. In an 
effort to evaluate these impacts, the Institute of Environmental Sciences (IES), 
Miami University, Oxford, Ohio, and the Whitewater Valley Jaycees, Union 
County, Indiana, cooperated on a social impact investigation. 

Traffic was monitored six times (twice each on holiday weekends, average 
weekends, and average weekdays) during the summer of 1976. Out-of-state vehicles 
ranged from 11.2% (regular weekday) to 23.7% (holiday weekend) of the total traf- 
fic volume on Indiana SR 101 and from 27.8% (regular weekday) to 35.5% (holiday 
weekend) on US 27. 

Questionnaires were distributed to visitors at the reservoir to determine the 
origin of people using this facility, the distance and routes traveled, and the 
economic and recreational activities in which they participated. A total of 957 ques- 
tionnaires was returned for analysis. Interestingly, only 42.2% of the respondents 
were residents of Indiana compared to 53.9% for Ohio. Only 1.3% of the visitors 
were from Union County, whereas 33.8% traveled through Union County to reach 
the reservoir. The highest frequency responded that they traveled 26 to 50 miles to 
reach the reservoir. Economic-wise, only 11.2% of these visitors made purchases in 
Union County. 

Residents of Union County were surveyed to determine the perception of and 

296 



Geology and Geography 297 

desires for the future of both the reservoir and Union County. Of the 800 question- 
naires mailed to residents, 203 (25.4%) were completed and returned. Expansion of 
local water facilities ranked as the most desirable need for change in the future. 
Loss of agricultural lands and loss of natural areas were ranked as the most 
undesirable changes. 

Engineering Geology of an Operating Area Strip Mine, Southwestern Indiana. 

Gerald E. Greengold and Terry R. West, Department of Geosciences, West 
Lafayette, Indiana 47907. Area strip mining of coal with concurrent reclama- 
tion (Pike County, Indiana) has involved some soil instability in spoil piles and ver- 
tical cuts. Overburden (50 to 180 feet thick) generally has a 1:3 to 1:6 ratio, soil to 
rock. Increases toward 1:2 yield spoil instability as rock volume is insufficient to re- 
tain the soil. 

Surface soils are ice-contact and glacial lacustrine sands, silts, and clays with 
extreme variability. Company-supplied drill logs plus borings for the authors, pro- 
vided data for plan maps and cross sections to correlate units and define their ex- 
tent. 

Stratigraphy is dominated by organic-bearing lacustrine clays and loose, fine 
to medium, saturated sands. Sands and clays range from small pockets to others > 
50' thick and hundreds of feet across. Larger deposits have an E-W elongation. 
Area partially covered by two distinct units ranging from 5 to 20 feet thick each: 
lower deposit-glacial drift, upper-windblown silt. Bedrock is gray, shale to sandy 
shale, interbedded with yellow and gray sandstone, and some limestone. The ir- 
regular bedrock surface presumably was formed by normal processes of erosion 
without glacial influence. Where soil overburden is thickest, sections contain more 
gray clay and/or saturated sand. 

Added borings had split spoon samples at 5' intervals to bedrock and some 
Shelby Tubes in zones thought to affect slope stability. Laboratory tests provided 
natural water contents and Atterberg limits. Shelby Tubes yielded natural den- 
sities and unconfined compressive strengths. Correlations between borings having 
spoons and Shelby Tubes with those of only split spoons allowed for characteriza- 
tion of materials. Clay strengths ranged widely from 0.04 to > 1.40 TSF. Sensitivi- 
ties averaged 1.7. Based on the stratigraphy and properties of the materials, poten- 
tial problem areas were delineated. 

Recommendations being considered for slope stability: removal of some soils 
in thickest sections prior to production excavation, dewatering of sands where ap- 
plicable and benching in thick soils to reduce cut-slope failures and to allow greater 
selectivity for soil-type placement in spoil piles. 

Groundwater Chemistry in Vigo County, Indiana. Joseph G. Hailer. Indiana 

Geological Survey, Bloomington, Indiana 47405. As part of an environmental 

geologic study of Vigo County, Indiana, 108 groundwater samples were collected 
and analyzed for major, minor, and trace element composition. Water from wells 
developed in the Pennsylvania-age sandstone aquifers of the eastern and north- 
western sections of the county are highly alkaline NaHC0 3 water. Fluoride, car- 
bonate, and chloride are the major secondary constituents in this water. Water 
from the more productive, unconsolidated sands and gravels of the Wabash River 
Valley is a Ca-HC0 3 -S0 4 water having high correlations with magnesium, stron- 
tium, silica, and nitrate. The occurrence of nitrate is likely related to human habita- 
tion and agricultural activity. Most chemical constituents are the result of the in- 



298 Indiana Academy of Science 

teraction with aquifer minerals. The thermodynamic equilibrium model, WATEQ, 
is used to describe mineral stability based on chemical composition of water. 

The Sedimentation of Lumsden Pond, Vigo County, Indiana. Eric Koglin, Depart- 
ment of Water and Natural Resources, South Dakota Geological Survey, Science 
Center, University, Vermillion, South Dakota 57069 and Donald W. Ash, Depart- 
ment of Geography and Geology, Indiana State University, Terre Haute, Indiana 

47809. Lumsden pond is located in a 10 acre drainage basin underlain by eolian 

derived soils. By determining the sediment accumulation over the pond's deposi- 
tional history, calculations were made to estimate the yearly rate of sediment 
yield. The east-west trending pond had ten north-south traverse lines plotted at 
equal distances along one side. Along each line, stations were established to record 
water depth and sediment thickness. Sediment thickness was determined by press- 
ing a steel probe into the sediment until a density change was noted. Two maps 
were plotted, one of the original bottom and another of the present bottom. Two- 
foot contour intervals were then drawn of the sediment thickness. Each contour 
was planimetered and the resulting data was used in the Modified Dobson 
Prismoidel Formula. The final result indicates that the average yearly sediment 
yield is 0.053 acre-feet. From this figure it was determined that the pond had lost 
1.06 acre-feet of storage over its twenty-year depositional history. By comparing 
these results with other surveyed ponds it was observed that accumulation in 
Lumsden Pond can be considered normal. Assuming nothing is done to modify the 
rate of sedimentation, the life expectancy of the pond is 82 years. 

Outlook for Shale-Oil Recovery from New Albany Strata in Indiana. John B. Pat 
ton and Richard K. Lenininger, Indiana Geological Survey, Bloomington, Indiana 

47405. Within the upper half of the New Albany Shale of southeastern Indiana, 

at least for sections so far analyzed, an interval of 30 to 40 feet contains an average 
of more than 10 percent organic carbon. According to data published concerning 
similar material, this interval should yield more than 20 gallons of shale oil per ton 
of rock when processed by hydrogen retorting. Large tonnages of the material are 
present at or near the surface, especially in Scott and Jennings Counties, and 
possibly Clark County, Indiana, and additional resources are accessible to under- 
ground mining where rock overburden is thick. Satisfactory reclamation of mined 
land, and other environmental measures such as protecting surface water quality, 
appear to be feasible. Ventures in shale oil recovery, with or without byproducts, 
are likely to be undertaken soon; and only the practical test of production at com- 
mercial level can establish the economic feasibility. 



Rurbanization and the Countryside Urban Web in Indiana 

Thomas Frank Barton 
Indiana University, Bloomington, Indiana 47405 



In contrast with urbanization, whereby an increasing proportion of the popula- 
tion in a political unit lives in settlements with populations of 2500 or more, rur- 
banization occurs when a higher proportion of the population in political units live 
outside of city boundaries, but do not qualify as farmers. Starting with the 1920 
Census the rural population has been divided into the two categories: nonfarm and 
farm. In 1970 the nonfarm family was rural annually selling less than $50 worth of 
crops or livestock from an area of 10 acres or more, or less than $250 worth from an 
area of less than 10 acres. If the income from crops and livestock sold was higher 
than the amount just described, the land was called a farm and the occupants were 
farm dwellers. 

Indiana's Rurbanization 

In 1920, rural nonfarm occupants accounted for 19 percent of Indiana's total 
population. The percentage increased to 28 by 1970. Percentagewise the 50-year 
growth of the nonfarm population does not appear too significant because urban 
growth was much greater. 

However, if one concentrates on the rural population only and compares the 
steep decline of the farm with the rapid growth of the nonfarm, the significance of 
this new category stands out in bold relief. During the 50 years 1920-1970, the rural 
farm population in Indiana declined from 902,820 to 372,862 — a decline of 242 per- 
cent. In contrast, during the same half century the nonfarm dwellers increased 
from 544,715 to 1,437,070 -an increase' of 273 percent. 

Countywise Rurbanization 

During the last half century, especially since World War II, countywise rur- 
banization has flourished. The number of counties in which the nonfarm population 
outnumbered that of urban and farm combined increased from nine in 1950 to 27 in 
1960 and 32 in 1970. The nonfarm outnumbered the combined urban and farm 
population in more than one-third of Indiana's 92 counties (Figure 1). Also by 1970 
rurbanization had developed so rapidly that, in addition to the 32 counties in which 
a majority were nonfarm dwellers, there were another 30 in which the nonfarm 
population was larger than the largest city in each county (Figure 1). County rur- 
banization reached a peak in Brown County where 89 percent of the people were 
nonfarm dwellers. In contrast, in 1960 only one county, Switzerland, located in 
southeastern Indiana, had over 50 percent of its population living on farms. 
However by 1970 over half of Switzerland's population was nonfarm. In 1970 only 
in 12 counties did the farm population account for 25-49 percent of the population 
(Figure 2). 

Distribution within Counties 

Within counties, rural nonfarm dwellers are found in four general areas. 
These are: (1) on one or both sides of all-weather roads, forming ribbon-like 
streamers or shoestring settlements extending for miles from a city boundary, 
often as far as the adjacent city or county seat in the next county, (2) in settlements 
which have populations of less than 2,500 (3) on the fringes of cities adjacent to city 

299 



300 



Indiana Academy of Science 




TFB/jmh 



Over half of the population are nonfarm dwellers 



H Nonfarm dwellers outnumber inhabitants of largest city 



Figure 1. Rural Nonfarm Dwellers in Indiana, 1970. 



Geology and Geography 



301 



xxj f- 1 I I | WABA H :-:.in! ixx ■ :'!: : 

^^CWJ WH,TE , i-, CASS JM.AM.j j ,0N |^|$$g 




VvOOOsV-^XxV^ STARKE I 

•;•■ :/. ■•■;::) jasper j^.&*£..y;3 



:•:;: ■:. ."v 




( parke I Ihendr-icks"; maf 

I I PUTNA" l\W 



I rush !fayette f: ;'/. ';-;'. 

H ELBY | U'.V .W/.^ '^^ 

| fe-XF r'an k'l IN!; 

vTdecatur S^*^**^ 




^ 21,000 - 42,000 Rural Nonfarm Population 
Hi 25 - 49 Percent Farm Population 



FIGURE 2. Farm and Rural Nonfarm Population Concentrations, 1970. 



302 Indiana Academy of Science 

boundaries and/or in leapfrogging subdivisions some distance from the boundaries 
and (4) although fewer in numbers, nonfarm dwellers also are found scattered about 
in a buckshot pattern throughout counties. Regardless of location, the nonfarm 
population has created numerous problems for the farm and urban people as well as 
for themselves. This paper will be limited to a treatment of the shoestring set- 
tlements which form the countryside urban web. 

Countryside Urban Web 

In contrast with the urban web of city structure, consisting of circulation- 
business-industrial corridors which enmesh residential and recreational land in 
cities, the corridors built in the countryside consist of all categories of urban land 
use although most of the land is used for residential purposes. These urban land 
use corridors enmesh the farm land. 

Prior to the 20th century in Indiana there was a comparatively distinct 
cleavage between land used for various city purposes and the surrounding farm 
land. But in the last half century, with the development and use of gasoline- 
powered vehicles and the construction of hard surface roads connecting the cities, 
the nonfarm dwellers began to buy land for residential purposes adjacent to all- 
weather roads serving the same purpose as paved streets in the city. Later, 
business establishments were added within the ribbons of nonfarm residences and 
then small and large industries located in these urban land-use corridors. Nonfarm 
uses of land multiplied and the corridor tenacles extending from cities grew in 
length until sometimes they coalesced with the corridors extending outward from 
adjacent cities. This growth produced a web of urban land-use corridors which sur- 
rounds the farm land. Generally, the meshes of the web are smaller near the cities 
and larger and disconnected midway between cities. The corridors are generally 
wider near the cities and decrease (but not uniformly) in width or become discon- 
nected between the larger settlements. 

In these urban countryside corridors, in addition to the all-weather roads, 
there are concentrated other circulation systems such as rural water, primary 
telephone and electric lines and truck, school bus and workers traffic routes. 

Some agriculturalists, urbanologists, government officials and planners, as 
well as members of the general public, believe these corridors are one of the forces 
most destructive of land, water, wildlife and scenic beauty to appear in the last half 
of this century. Traffic flows are increased by vehicles transporting nonfarm 
dwellers to cities. Stop signs and stop lights are placed at closer intervals on the 
primary roads, slowing traffic speed and leading to congestion. The situation just 
described is widespread in the countryside corridors of Marion County. 

By living in the county the nonfarm dwellers must drive long distances weekly 
to the city to work, to shop and for social engagements. This extra mileage results 
in an enormous waste of energy, time and natural resources used in powering and 
replacement of vehicles and maintenance of highways. 

These urban land-use corridors create additional problems of (1) natural 
drainage distribution, which leads to flooding, (2) inadequate human and other 
waste disposal, (3) acceleration of pollution of various water resources by failing 
septic fields, (4) proliferation of poor tertiary roads, (5) illegal disposal of garbage 
and trash, (6) disruption and blocking of wise land use and proliferation of poor land 
use, (7) skyrocketing public service costs and (8) spawning of rural slums. 



Geology and Geography 303 

Decade of the 1970s and 1980s 

Will the 1980 census reveal an increase, decline or leveling off of the growth of 
rurbanization in the 1970s in Indiana? Will rurbanization continue throughout the 
1980s? Some of the variables which need to be considered in making projections 
are: (1) definition of a farm, (2) speed of annexation, (3) county metropolitan govern- 
ment, (4) enforcement of better land use, (5) transporation costs, (6) increase in 
public mass transit and (7) changes in technology. 

Definition of a farm. One of the primary reasons for believing that rurbanization 
continued during the 1970s is that in 1975 the U.S. Department of Agriculture and 
the Census Bureau changed the criteria used to classify an acreage as a farm. In the 
1980 census, for an acreage to be classified as a farm, the owner and/or renter must 
sell annually at least $1,000 worth of livestock or crops, regardless of acreage size. 
Before this change one could only qualify as a farmer if his annual sales amounted 
to $50 from 10 acres or more, or $250 from less than 10 acres. The size of the 
acreage is no longer a criteria in the new definition. Although in 1976 it was 
estimated that the new definition would declassify over half a million farms in the 
United States, a decrease of 16 to 20 percent, accelerated inflation in the last five 
years may have greatly reduced these estimates. Many believe that the 1975 
criterion was not too realistic and that inflation has played havoc with the dollar 
amount of sales. It takes only one of a few horses, cattle, pigs, sheep or even dogs to 
bring $1,000. 

Annexation. If the city annexation process were not so handicapped in Indiana by 
excessive requirements established in court cases and by long drawn out court bat- 
tles, annexation could turn a large number of nonfarm dwellers (clustering adjacent 
to but just outside city boundaries) into urban dwellers. Annexation has moved at a 
snail's pace in the 1970s, and there is little likelihood that annexation will greatly 
reduce the rurbanization process in the next decade. 

County metropolitan government. The adoption of county metropolitan govern- 
ment eliminates the nonfarm dwellers statistically but not the problems they have 
created. Moreover only Marion County (where the state capital is located in In- 
dianapolis) has adopted Unigov. It seems improbable that any other county will 
adopt this form of government during the next decade, so county metropolitan 
government soon may not be a deterrent to rurbanization. 

Better land use. In many of the 62 counties in Indiana where the nonfarm dwellers 
outnumber the urban and farm population or outnumber the dwellers in the largest 
city, county or area planning agencies are so weak and ineffective that nonfarm 
residential use has a higher priority than farming. In only a few of these counties 
have post World War II soil surveys been made and published that contain 
valuable information on which to base wise land use decisions. Even in the counties 
where recent soil surveys have been made, the nonfarm population has grown so 
large, and carries so much political clout, that it is very difficult to bring about wise 
land use reform. Apparently, most nonfarm dwellers are not adequately informed 
on the principles of management of land, water, vegetation, wildlife and geologic 
resources. Consequently too often they flock to public hearings and engage in quar- 
relsome, negative debate. They interrupt and shout down those who attempt to 
present reasonable considerations. The road leading to attainment of wise land use 
appears to be a long rocky one in counties dominated by nonfarm dwellers. Wise 
land use policy is at present an insignificant deterrent to rurbanization in Indiana. 



304 Indiana Academy of Science 

Transportation costs. In the 1980s rurbanization may be slowed down, halted or 
reversed if gasoline costs soar to over $2.00 per gallon and automobile and truck 
prices continue to climb. Most nonfarm families of four with two pre-college age 
children, and with both husband and wife working, have two or more cars. Many of 
these nonfarm commuters drive hundreds of miles each week. The school bus may 
pick up their children and take them to the consolidated schools, but the parents 
still need to drive children under 16 years to many school, church, city and other ac- 
tivities. People who decided to buy an acreage and build a home in the country five 
to 10 miles or more from the city prior to and since 1973 when gasoline was fifty 
cents a gallon, may plan to move to the city when gasoline reaches $2.00 or more. 
People who were attracted to country living because taxes were lower may soon 
realize that part or most of the tax advantage has been wiped out by the soaring 
costs of driving a car or truck. 

In addition to these costs of commuting, the nonfarm population faces the 
possibility of road deterioration due to skyrocketing costs of road maintenance and 
the problem of securing enough funds to maintain the present roads. Moreover the 
expansion of the all-weather roads may grind to a halt. The corridor dwellers have 
not only put a heavy burden on road maintenance but also they have exerted heavy 
political pressure for additional all-weather roads. 

Public mass transit. The growth of city mass transit may become attractive enough 
to influence some nonfarm dwellers in a few counties to come back to the cities and 
encourage others not to buy or build residences in the country-side. However, 
public mass transit will not exert too great a pull on these dwellers in many coun- 
ties because the cities in many of the counties may be too small to develop public 
mass transit. In 1970 Indiana had only nine cities with populations of over 50,000 
but 142 cities with populations of less than 50,000. These smaller cities are concen- 
trated in the same counties as the nonfarm population. 

Indiana has taken action to develop public mass transit in its cities. In 1980 the 
Indiana legislature passed a mass transit funding bill that will probably provide at 
least $10 million to help develop and operate public transit systems in 1981. 
Transportation was granted 1 percent of the state gross sales tax revenues and 
mass transit is to receive 95 percent of that. In the past public mass transit had 
been handicapped by inadequate local funding. In 1979 the state was allocated 
about $25 million of federal funds for mass transit but about $15 million of it was 
never used, because local funding was not available to provide matching funds. 

Although city mass transit is expected to grow in Indiana, the nonfarm 
residences are so widely scattered that county-wide mass transit in the country- 
side urban corridors should not be established because it would necessitate 
unreasonable subsidization. 

Changes in technology and economy. Unfortunately one cannot foresee the 
changes in technology or the economy or political policy that may stimulate the 
growth or decline of rurbanization. Many such diverse factors as (1) power lawn 
mowers and garden tools enabling people to more easily care for large residential 
areas, (2) rapid expansion of all-weather roads, (3) the rapid growth of 4-H programs 
requiring larger areas for raising livestock and (4) federal policy of subsidizing 
rural electrification, rural water systems and rural development in general, have 
stimulated rurbanization both in Indiana and other states. 

Summary 

The percentage of nonfarm dwellers in the total population of Indiana increased 



Geology and Geography 305 

from 19 percent in 1920 to only 28 in 1970. During the 1960s rurbanization leveled 
off and grew less than one percent. But in 1970 the nonfarm population numerically 
exceeded that of Indiana's nine largest cities with populations of over 50,000, or the 
remaining 142 cities. Rurbanization saps the population growth of cities and 
creates a web of urban land use that surrounds farm land and disrupts proper use 
of natural resources. The web pattern of urban land use corridors interferes with 
the growth of both agriculture and cities — and is one of the most wasteful types of 
urban land use. In spite of stimulating the growth of rurbanization by subsidiza- 
tion, the federal government in 1974 published a volume entitled The Costs of 
Sprawl- Environmental Costs of Alternative Residential Patterns at the Urban 
Fringe. This volume stresses the fact that nonfarm development on the fringes of 
cities is the most expensive method of community development. The study was 
made by a private company. The volume was published by the Superintendent of 
Documents, U.S. Government Printing Office, Washington D.C. 20402. 

Now we may need a private study of the urban web in the countryside sub- 
sidized by the federal government. Until such action is taken, geographers may 
find rurbanization and the countryside urban web interesting topics for research. 
County planners and county commissioners as well as other appointed and elected 
county administrators desperately need information which can be provided by 
county and regional studies. These studies could provide valuable data needed for 
planning and regulating urban land use development in the countryside corridors 
that are already in existence, and regulating more carefully future rurbanization of 
all kinds. More information is needed before administrators can carefully decide as 
to whether to make illegal, reduce, restrict or plan and zone corridor rurbanization 
growth. 

Will geographers and others in the various university disciplines provide the 
studies needed? 



A Statistical Correlation of Sulfur Content in Coal 5 
and the Overlying Gray Shale 

S. N. Ghose 
Amax Coal Company, Indianapolis, Indiana 46225 

Introduction 

The marketability of coal today largely depends on sulfur content - ash and 
BTU being comparable. Low sulfur coal, which is not abundant in the Midwest, is in 
great demand and to an extent, may significantly affect the future of the coal in- 
dustry in that region. Competition for low sulfur coal has influenced the 
midwestern coal industry from exploration to operation. 

The association of relatively low sulfur coal with overlying thick, freshwater 
gray shale is a familiar fact in the geology of the Illinois Basin. This interesting 
phenomenon is mentioned in the literature by various authors such as Hopkins, 
Wier, Friedman, Eggart and others. Hopkins (1968) described this association in 
detail and the function of the gray shale in preventing exterior sulfur contamina- 
tion of the coal. In this paper, an effort is made to establish an empirical relation 
between these two variables: gray shale thickness and total sulfur content. Based 
on these variables a predictive scale is constructed to approximate the total sulfur 
content per given gray shale thickness for quick appraisal. This correlation is based 
on the data of Coal 5 which is a major commercial seam in the Illinois Basin. Coal 5 
shows significant uniformity in quality, thickness and roof lithology. Also, unlike 
Coal 6, Coal 5 does not show any persistent parting (blue band). Therefore, the 
depositional environment of Coal 5 is probably less complicated and hence is easier 
to correlate and predict the total sulfur content. However, the method presented 
here also can be applied to other coal seams for predicting total sulfur content 
under similar geological conditions. 

Coal 5 in the Illinois Basin normally shows two distinctive roof lithologies: ex- 
tensive marine black shale and local fresh water gray shale. Although several 
geological conditions can lead to the deposition of a fresh water gray shale, the 
gray shale encountered in Coal 5, in many parts of the basin, is that of an active 
channel origin. Periodic flooding of the coal swamp near the channel deposited the 
fine sediments on Coal 5 near the channel as shown in Figure 1 and Figure 2. 

Procedure 

In this study ninety analyses of Coal 5 core samples and the corresponding 
thickness of overlying gray shale are used as the data base for defining the attitude 
of the curve shown in Figure 3. The characteristics of these two variables are cor- 
related and categorized at 5 feet intervals. Each 5 feet of increment (unit block) pro- 
vides one mean position on the curve. Presumably, the smaller the unit block, the 
greater expression of curve-detail which in turn adds to the accuracy of the 
method. As shown in Figure 2, the degree of scatter is considerable, and the 
overlapping of data is unavoidable if point to point correlation is desired. The scat- 
ter of data points represents variations in the depositional system. Therefore, to 
obtain a meaningful interpretation within the limits of practicality, 5 feet unit 
blocks were chosen. To provide a measure of reliability the basic statistical 
parameters of error distribution are estimated for each incremental 5 feet (Table 1 
and Table 2). The data represented in each block are free from systematic error 

306 



Geology and Geography 



307 



POSSIBLE THICK COAL AREA 
WITH GRAY SHALE TOP 
(GOOD MINABILITY) 



ANASTOMOSING 
CHANNEL 



NATURAL LEVEE 




Figure 1. Diagrammatic representation of Sandstone Channel with relation to 
Coal V Swamp Environment 



because Evi = 0. 

i = l 



Sample means from each 5 feet interval block were fitted by computer. The 
best fit is a polynomial regression of degree two. The observed and estimated 
sulfur percentages are given below. 



Gray Shale 


Observed 


Estimated 


Thickness 


(Percent) 


(Percent) 


5 


3.9860 


3.6465 


10 


3.9600 


3.1096 


15 


1.8575 


2.6288 


20 


1.5360 


2.2042 


25 


1.5162 


1.8358 


30 


1.3850 


1.5235 


35 


1.2750 


1.2674 


40 


1.1175 


1.0674 


45 


1.3375 


.92350 


50 


0.7100 


.83579 


55 


1.1175 


.80422 


60 


1.3855 


.82880 


65 


.9340 


.90953 


75 


.70667 


1.2394 



To make this correlation easily usable and to avoid overlapping errors further 
adjustment and refinement is necessary. This refinement is achieved by expanding 
the incremental gray shale unit or condensing the predictive scale. A 4 step scale is 
proposed here, which is more compatible with the margin of error and allows the 
steps to be more easily identified. 



308 



Indiana Academy of Science 



CHANNEL 
(Coal Absent) 




BLACK SHALE 
(MARINE) 



BLACK SHALE 
(FRESH-WATER) 



Figure 2. Schematic cross-section relating environments of deposition to coal 

and roof lithologies 



Black Shale 

2 to 10 feet of gray shale 
10 to 25 feet of gray shale 
25 to 80 feet of gray shale 



3.5% to 6% or more sulfur 
2% to 3.5% sulfur 
1.5% to 2% sulfur 
0.5% to 1.5% sulfur 
Analysis 



• BOREHOLE SAMPLES 
o SAMPLE MEAN 
A RANGE 




20 25 30 35 40 45 50 55 

GRAY SHALE THICKNESS Ft. 



Figure 3. Correlation of Total Sulfur Percentage & Gray Shale Thickness 



Geology and Geography 



309 



Of CM 



O 1ft 



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3 



£ 3. S $5 



« E ._ <^._ 5 £ x £ x> 

U * > > > Z <d £ «> 



5 £# 



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t £ s |a 1 1 i >< 



310 



Indiana Academy of Science 



*- 



*l - 



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Geology and Geography 311 

A comparative study of statistics to test the relation between the base and the 
predicted quantities and homogeneity of the population indicates moderate agree- 
ment. 

The covariance shows that there is a positive correlation (both increasing) 

covariance = Q xy = + 0.042772 

Icxyl < le x e y l 

0.042772 0.116227 

The covariance is smaller than the standard error of the two sets combined which 
indicates that these two sets are in good agreement. 

The correlation coefficient (Pxy) between these two sets is 0.3680007 which is 
moderate. This suggests that other variables such as seam thickness and distance 
from the cut out affect the correlation. 

An F test was given to check the equality of variances of these two sets. In 

other words hypotheses, Ho: Q* = £f was tested against 

Q*± ef at the 
significance level oc = .10 (1-oc = confidence level). The degrees of freedom for 
both sets are taken as 8. 







F = 


8J 

k 


" = 


0.3947540 










oc = 


.10, 


la 


12 = 0.95, 


a/2 = 


.05 


F (1- 

r DF 1 DF 2 u 


a/2) = 




1 








F DF 2 DF ] 


(a/2) 




From the table, 


















F 8,8,0.95 " 




1 


1 

^ 9 AA 


0.2907 






Fo 


q n a 





0.39 > 0.29 do not reject the hypothesis 
0.39 < 3.44 do not reject the hypothesis 



Conclusion 

The method described in this paper, can be used not only in the regional type 
of study but also as an aid to select the most viable prospect among many for ex- 
ploration. Also, in the developmental stages the method can be utilized to selective- 
ly increase the density of information. Further study of the homogeneity of the 
population, and analysis of data from different seams may provide a quantitative 
base from which environments of deposition can be assessed in order to locate good 
quality coal. 

Acknowledgements 

I am thankful to Dr. C. E. Wier for his permission to publish this paper. Mr. D. 
A. Darko and Mr. T. R. McCarthy painstakingly reviewed the manuscript and of- 
fered a number of useful comments for which I am grateful. I also wish to thank Dr. 
K. Badiozamani for his help in curve fitting by the computer. 



312 Indiana Academy of Science 

Literature Cited 

1. Bevington, P. R. 1969. Data Reduction and Error Analysis for the Physical 
Sciences: McGraw-Hill Book Company, N.Y. 

2. Eggart, D. L. and S. C. Adams. 1979. Distribution of Fluvial Channel Systems 
Contemporaneous with the Springfield Coal Member (Middle Pennsylvanian) 
in Southwestern Indiana: Ninth International Congress of Carboniferous 
Stratigraphy and Geology, University of Illinois, Urbana-Champaign. 

3. Friedman, S. A. 1955. Split and Channel Sandstone Cut Out in Coal V in the 
Dresser Area, Vigo County, Indiana: Indiana Academy of Science, v. 65, P 
165-168. 

4. Hopkins, M. E. 1968. Harrisburg (No. 5) Coal Reserves of Southern Illinois; Il- 
linois Geol. Surv. Circ. 431. 

5. Pugh, E. M. and G. H. Winslow. 1966. The Analysis of Physical 
Measurements: Addison-Wesley, Reading, Massachusetts. 

6. Wier, C. E. 1952. Distribution, Structure and Mined Areas of Coal in Sullivan 
County, Indiana: Indiana Geological Survey Prelim. Coal Map 2. Wier, C. E., 
and Hutchison, H. C, 1977, Reduction of Sulfur in Indiana Coal by Washability 
Techniques: Department of Natural Resources, Geological Survey Bulletin 55. 



An Analysis of Irregularity of Surveyed Sections in Indiana 

Andrea Sjoreen 

Indiana Geological Survey, Bloomington, Indiana 47405 

Richard L. Powell 

Geosciences Research Associates, Inc., Bloomington, Indiana 47401 

Introduction 

The Congressional Land Survey System, commonly called either the Section, 
Township, and Range or the rectangular land system, is frequently used to make 
accurate descriptions of geographic location. The extent to which sections are not 
square and do not have sides of exactly one mile can affect the complexity and ac- 
curacy of the location description. It was possible to measure the lengths of all sec- 
tion sides in Indiana to an accuracy of about ± 25 feet using data collected for a 
computer mapping system. Using the length of each section side made it possible to 
assign a degree of irregularity to each section in the state. The computer mapping 
system, coupled with a Calcomp plotter, has drawn maps locating the sections with 
varying degrees of irregularity as well as a map of the most nearly perfect sections. 
The maps illustrate some of the systematic error in the original land surveys and 
point out some errors or differences that are not easily explained. 

The locations of all the section corners in Indiana were digitized by the staffs 
of the Illinois and Indiana Geological Surveys from IVi minute topograhic 
quadrangles published by the United States Geological Survey. The computer map- 
ping system was designed at the Illinois Geological Survey. This system is design- 
ed to handle sections only and not locations, military donations, surveys, reserva- 
tions, or other non-Congressional surveying units. All of these extraneous units are 
here considered irregular, and were not considered pertinent to the current study. 
Their presence did not significantly affect the results of this study. The analysis of 
section irregularity was conducted at the Indiana Geological Survey with the en- 
couragement and assistance of Dr. Robert F. Blakely. 

The Congressional Land System 

The Congressional Land System was enacted by the Continental Congress on 
May 20, 1785. Townships and sections were to be surveyed on the ground with the 
accompanying legal records of land ownership for most of the usable lands west of 
the Allegheny Mountains. Several schemes of systematic rectangular surveys 
were attempted in the State of Ohio. These prototype surveys were more useful 
than the highly irregular metes and bounds system of land survey used in Ohio and 
other states at that time. The present plan of townships, 6 miles to a side, subdivid- 
ed into 36 sections, each one mile square, was adopted in western Ohio and 
southwestern Indiana by an Act of Congress on May 18, 1796 (3). Other minor in- 
structions governing the Congressional Land Survey System were enacted by Con- 
gress on March 26, 1804. 

The first lands surveyed by any system in Indiana were probably the French 
farmlands in the vicinity of Vincennes. The first rectangular survey in Indiana, 
although a non-Congressional plan, was the Illinois Grant, also called Clark's Grant, 
mostly in Clark County. The lands were donated by the Virginia General Assembly 
in 1781 to General George Rogers Clark and his regiment and surveyed by William 
Clark in 1795 or 1796. Other Military Donation Lands were established in Knox 

313 



314 Indiana Academy of Science 

County by Congress in 1891. Other lands in the vicinity were surveyed as Loca- 
tions and Surveys to relocate those disposed from the Military Donations (2). 

Various survey lines were established in the Northwest Territory and later 
Indiana between 1802 and about 1846, some into adjacent states, to establish the 
boundaries of a series of concessions of lands resulting from treaties with various 
Indians. About 20 separate treaties were made for lands now in Indiana and some 
irregular tracts remain as Indian Reserves. 

The first Congressional Land System survey in Indiana was initiated in 
southwestern Ohio as a result of the Greenville Treaty of 1795. Lands were 
surveyed as townships east and west of a meridian extending north from the mouth 
of the Miami River, later established as the First Principal Meridian, and the 
Ohio — Indiana State Line in 1810. A somewhat triangular area was laid out in 
townships and sections from 1799 to 1805. Treaties at Fort Wayne in 1803, Vincen- 
nes in 1804, and Grouseland in 1805 acquired the southern fourth of the present 
state. 

The initial point for the Second Principal Meridian and Base Line was 
established astronomically by Jared Mansfield in 1804, within about two miles of 
Latitude 38° 30' North and Longitude 86° 30' West. Ebenezer Buckingham 
surveyed the Base Line in 1804 and subsequently lands within the Vincennes Tract 
were surveyed in accordance with the Congressional System from 1804 to 1806, ex- 
cepting the Military Donations, Locations, Surveys, etc. previously granted by 
Congress. The township and section surveys proceeded southward in 1805 to 1806, 
eastward in 1806, then northward to the Michigan line by about 1829-1830, generally 
soon after each land concession by Indians (1). A few Indian concessions were not 
surveyed until as late as 1840. 

Township and Section Surveying Procedure 

Simply stated, range lines were laid out as true north-south meridians at six 
mile intervals east and west of the Second Principal Meridian and township lines 
were established as parallels extending east-west at right angles to the range lines 
at six mile intervals from the base line. Section and quarter section monuments 
were set at half mile intervals along these lines as they were surveyed. The range 
lines noticeably converge poleward such that standard parallels or correction lines 
were established as new base lines every 24 miles or every four townships, starting 
anew with a full 24 mile width from each correction line. 

The survey of mile square sections with quarter section markers within each 
township was to proceed from the southeast corner of the township. The eastern 
most row of sections was surveyed first, from south to north, then the next row of 
sections on the west, from south to north, to establish five rows of sections. The 
westernmost row was surveyed last. Each section was intended to contain 640 
acres, but the Land Office was aware of the imperfections of surveying, as well as 
the faults of some surveyors, perhaps, and expected some errors in addition to 
those caused by convergence of the meridians. Error from south to north was to be 
reconciled within the northern half section of the northern tier of sections (Sections 
1 through 6). Deviation from a perfect section east to west, particularly that caused 
by convergence, was to be within the western half of the western row of sections 
(Sections 6, 7, 18, 19, 30, and 31). Sections within these two rows that contain ap- 
preciably more or less than 640 acres are called fractional sections. Theoretically, 
sections not in these two rows should be perfect sections of 640 acres each. Such is 
not the case. 



Geology and Geography 



315 



Anyone with even limited experience in determining exact locations using 
footages within the quarter, quarter, quarter parts of conventional sections quickly 
becomes aware of the minor imperfections within the system. The exact distribu- 
tion and amount of deviation of fractional sections has until now been unavailable, 
although many such errors had been noted, such as slivers of some sections less 
than a quarter mile wide north-south (northeastern Posey County) and elongated 
sections more than a mile and a half long (southeastern Washington County). In- 
itially, this study was based on the assumption that of the 36 sections in a township 
25 should be essentially perfect and that 11 were fractional sections with most of 
the error occuring in the northern tier and western row of sections. The amount of 
deviation of the fractional sections was to be used as a measure of the imperfection 
of any so-called perfect sections, or, overall, a maximum of about 30 percent of the 
sections should be considered irregular, the remaining 70 percent, more or less, 
should be regular by definition, or at least up to federal standards. 

Method of Analysis 

All the section corners in Indiana were digitized from standard l x k minute 
topographic maps in longitude and latitude using a Summagraphics digitizer. The 
digitizing equipment was accurate to 0.005 inches. The determination of section 
corner location could be made accurate to half the mapped road width which is 
about 25 feet on the maps used. Section side length was calculated as the arc length 
between the appropriate section corners. Where sections were bounded by a river, 
the river side of the section was calculated as the arc distance between the end- 
points and not as the length of the river between them (Figure le). 

The degree of irregularity of each section was calculated as the maximum ab- 
solute difference of any of its sides from 1.00 mile. The calculated irregularities of 
some sections are shown in Figure 1. Sections which were so irregular as to have 



1.00 




1.00 1.2 5 



LOO 



£5 



.25 



.00 



.6 



99 



LOO 

Figure 1. Schematic representation of possible irregularities of sections. Sec- 
tions (a), (b), and (c) are nearly square. Sections (dl and if) have been terminated 
along an Indian treaty line. Section (e) has one side along a river on a state boun- 
dary. 



316 



Indiana Academy of Science 



only three sides have an irregularity of 1.00 mile (Figure Id). For this analysis, sec- 
tion lines are considered to cross county lines, but not to cross state lines. 

The deviation of each side from true north-south or east-west was also com- 
puted. This irregularity was computed as the difference in latitude of the ap- 
propriate section corners for east-west section lines and as the difference in 
longitude for north-south lines. Sections which were irregular due to an angular ir- 
regularity always were irregular due to a length irregularity. So to simplify the 
analysis, only the irregularity in length was used to assign the degree of irregularity 
to each section. 

Results 

A histogram showing the distribution of actual section irregularity is shown in 
Figure 2. This distribution has a mean of 0.03 miles, a median of 0.04 miles, a mode 



mperf ections 



mode 
(.01) 



6000 



5000 



— 4000 



median 
(.007 ) 




— 3000 



— 2000 



20 miles 



Figure 2. Histogram of irregularity for 36,698 sections in Indiana, This is the 
maximum difference of any section side from 1.00 mile. 



Geology and Geography 



317 



of 0.01 miles, and a range of - 1.00 to 1.10 miles for a total of 36,678 sections. 
Because this is clearly not a normal distribution, no further statistical analysis 
would be meaningful nor would it be meaningful to try to establish a statistical 
basis for section irregularity. Figure 3 shows a histogram of the distribution of the 
section side lengths. This distribution has a mean of 0.99 miles, a median of 1.00 
miles, a mode of 1.00 miles, and a range of 0.00 to 2.81 miles. Again this is not a nor- 
mal distribution and no further statistical analysis would be meaningful. The sharp- 
ness of the peak of this distribution shows the accuracy of the section surveys. 
Sixty-two per cent of the section sides are correct to within .01 miles (53 feet). The 
percentage of sections accurate to this limit, 29 percent, is much lower because 
whole section accuracy is measured by its worst side. 

A criterion for section irregularity is suggested by the method by which the 
surveying was done. Because all townships were surveyed from southeast to north- 
west, most of the surveying error is expected to accumulate in the northern and 
western tiers of sections within each township. A township diagram showing the 
percentage of imperfect vs. perfect sections is shown in Figure 4. Most of the im- 
perfect sections are clearly in the northern and western tiers while most of the 



Section Line Lengths 

mode ( LOO) 
— 24,000 /median (1.002) 



20,000 



6,000 



— 12,000 



— 8,000 



— 4,000 



mean 
(.992 



\ 



— 



I I 1 r 

.80 .90 1.00 1. 10 miles 

Figure 3. Histogram of lengths of 85,316 section lines in Indiana. As expected the 
mode is 1.00 mile. 



318 Indiana Academy of Science 

30 % 34.0 
20% 41.4 
I07o 46.3 
Perfect Sections % 4.3 



30% 32.1 

20% 35.8 

10% 33.4 

Perfect Sections % 4.0 



r 6 




5 




4 


■ 


3 


r-" 


2 




i 










■ 




■ 




■ 






n 7 




8 




9 


10 


II 


12 




_[ 


ml 


rm 1 


rm 1 


rm 1 


rm 1 


18 




17 




16 


15 


14 


13 


■ 


_J 


Tnl 


_[tT] 1 


rm 


rm 1 


rrn 1 


19 




20 




21 


22 


23 


24 


■ 


r 


™i 


rhn 1 


rhn 1 


rm 1 


rm l 


30 

■ 


29 


28 

rm 1 


27 

rhn 1 


26 

rm I 


25 

rin 1 


31 




32 




33 


34 


35 


36 


■ 


J 


rn 1 


rm 1 


rr-h 1 


rm 1 


rm 1 



30 K\v De v i a t ion 
I0°/N Classes 



10 

a> 

Q. - 



Perfect 
Section 



Figure 4. Schematic representation of the sections in a township showing the 
percentages of imperfect and perfect sections by section number. 



perfect sections are not (59 percent of the imperfect sections vs. 8 percent of the 
perfect sections). The sections in these two tiers include 11 of the 36 sections in a 
township or 30.5 percent. Assuming 30 percent as the cutoff for irregularity, sec- 



Geology and Geography 



319 



tions which vary from 1.00 mile by 0.03 miles (158 feet) are considered irregular. A 
computer generated map showing the locations of this 30 percent is shown in 
Figure 5. 




Figure 5. Locations of the 30 percent worst sections in Indiana. These sections 
have at least one side varying from 1.00 mile by ± 0.035 mile (185 feet). 



320 Indiana Academy of Science 

Discussion 

The map in Figure 5 shows that irregular sections do tend to be on the north 
and west sides of townships, but that is not always the case. Other less obvious fac- 



r 



J ' J 



38 

37 — 
36 

35 
34 
33 

"\ 

30 j 

29 



26 



T7 



# 



m 



■ 

23 
22' 



T2 




Figure 6. Locations of the 10 percent worst sections in Indiana. These sections 
have at least one side varying from 1.00 mile by ± 0.10 mile (528 feet). 



Geology and Geography 



321 



tors such as the presence of wide rivers, rough terrain, or swamps certainly af- 
fected the accuracy of surveys. For comparison Figure 6 shows the worst 10 per- 




: ; r? ■ - ... " — ! — :: — i 

. . mm : ,.-. .-;.. 



VoOmiL hJtl b Vk 2 P l TCent 0fs f cti ° nS in Indian(L These W sections ™ryfi 
1.00 mile by less than the error of measurement, ± 25 feet. 



rom 



322 Indiana Academy of Science 

cent sections. These clearly have a much stronger tendency to be in the northern 
and western tiers of sections than on the 30 percent map. The map of "perfect" sec- 
tions (Figure 7) shows no particular pattern to the locations of these best sections. 
This map shows all the sections with all sides equal to 1.000 ±0.005 miles; 0.005 
miles being the inherent error in the section corner location measurement. The 
diagram in Figure 4 has shown them to not be in the northern and western tiers 
within each township. These sections are fairly evenly distributed across the state, 
with a tendency to be more common in the north than the south. This is probably 
due to the presence of the Knox and Clark county irregular surveys and the large 
number of wide rivers in the south. 

Any concept of perfection within the Section, Township and Range (Congres- 
sional Land) Survey System is unfounded inasmuch as there is an average of less 
than one perfect section per township in Indiana. This analysis for Indiana has 
shown that 30.7 percent of the sections have at least one side varying more than 3 
percent (158 feet), 20.7 percent varying more than 5 percent (264 feet), and 9.9 per- 
cent varying more than 10 percent (528 feet or .10 mile). Fractional sections on the 
western row and northern tier of sections account for 69 percent of the sections 
that vary more than .10 mile much as planned by Congress in 1796. Much of the re- 
maining 31 percent is accounted for by sections which terminate along the Ohio and 
Wabash Rivers and so cannot be whole sections. Yet some errors greater than .10 
mile as well as smaller yet significant errors occur in places where gross survey er- 
ror is their cause. 

Several reasons for various errors of greater or lessor magnitude appear ob- 
vious. Generally, the amount and number of errors, aside from the normal occur- 
rence of fractional sections, seem to diminish northward in Indiana as the surveys 
progressed with time. Assuming this to be true, the better surveys may be 
credited to: 1) flatter topography in a northerly direction, 2) development of better 
survey instruments, 3) better instructions, and 4) more quality control by the 
government. Some surveyors conducted relatively shoddy surveys. On the other 
hand, topographic features alone may account for many errors, even in northern In- 
diana, considering the non-traversable obstacles such as lakes, swamps, and some 
rivers present at the time of the surveys. 

References 

1. Collett, John 1883. United States surveys and growth of timber: Indiana 
Dept. Geology and Natural History, 12th Annual Report, p. 26-38. 

2. Schneider, A. F. 1965. Noncongressional land-survey divisions in Indiana: 
Proc. Indiana Academy Science, vol. 74, p. 248-254. 

3. Sherman, C. E. 1925. Original Ohio Land Subdivisions: Ohio Cooperative 
Topographic Survey, Vol. Ill, 233 p., 9 figs., 40 pis., 1 chart. 



Faulting In Perry and Spencer Counties, Indiana 

Dan M. Sullivan, Curtis H. Ault, and George F. Tanner 
Indiana Geological Survey, 
Bloomington, Indiana 47405. 

Introduction 

Perry and Spencer Counties border on the Ohio River in southwestern In- 
diana at the juncture of the Wabash Lowland and Crawford Upland physiographic 
provinces. Spencer County lies in the Wabash Lowland where the exposed bedrock 
formations of the county are composed of shales, clays, coals, sandstones, and thin 
limestones of the Pottsvillian and Alleghenian Series of the Pennsylvanian System. 
Pennsylvanian rocks that have been faulted in southern Spencer County are 
covered by alluvial deposits of the Ohio River and other Pleistocene to Holocene 
sediments. 

Perry County lies immediately east of Spencer County within the Crawford 
Upland. The erosion of alternating sandstones, shales, and limestones of the 
Chesterian Series (Mississippian) and sandstones and shales of the lower Potts- 
villian Series has resulted in rugged, angular topography, which facilitates surface 
mapping of the faults in the southern part of the county. 

Faulting in Perry County has been known for more than a century. Small 
faults in a coal-mine tunnel near Cannelton were mapped early by E. M. Kindle, 
who also noted a fault with 35 feet of throw in a bluff near Cannelton (1). Faulting 
near Deer Creek was noted by Joseph Lesley, Jr., before 1862 (11); later workers 
viewed this possible fault with some doubt until it was confirmed after 1900. Many 
small tunnel coal mines were opened near Cannelton beginning about 1835 (5), and 
the early miners encountered small faults in some mines (1). 

In 1951 Hughes (8) mapped faults in the Deer Creek area, and more recently 
the Indian Creek Fault was shown on a regional geologic map at a scale of 1/250,000 
by Gray, Wayne, and Wier (7). 

During the past several years geologists of the Indiana Geological Survey 
have conducted intensive investigations of the faults in all of southwestern Indiana. 
This study of the faults in Perry and Spencer Counties was part of a project funded 
in part by the Nuclear Regulatory Commission to study the tectonic processes 
within a 200-mile radius of the active seismic center of New Madrid, Missouri. 

Tectonic Setting 

Perry and Spencer Counties lie on the eastern flank of the Illinois Basin, which 
is bounded on its northeast side in Indiana by the Cincinnati Arch. 

The faults in Perry and Spencer Counties are 50 to 80 miles east of the more 
extensive Wabash Valley Fault System, which is well developed in southwestern 
Indiana in Posey and Gibson Counties (2, 13, 14). 

The Wabash Valley FaultJSystem, which includes faulting in northwestern 
Kentucky and southeastern Illinois, trends north-northeastward, as do the faults in 
Perry and Spencer Counties. Three of the faults in Perry and Spencer Counties are 
extensions of faults mapped in Kentucky by Goudarzi and Smith (6), Johnson and 
Smith (9), Bergendahl (3), and Mayfield and Chisholm (10). 

The vertical displacements of the rocks faulted in Perry and Spencer Counties 

323 



324 Indiana Academy of Science 

are considerably less than those of the major faults in the Wabash Valley Fault 
System. All known faults in both areas are of the high-angle normal type. Although 
faulted rocks at the bedrock surface in Perry and Spencer Counties are older than 
those in the Wabash Valley area, the time of faulting of the Pennsylvanian beds in 
Perry County probably relates closely to the post-Pennsylvanian and pre- 
Pleistocene time of faulting of the Wabash Valley Fault System. 

The relationship of the Perry County and Spencer County faulting and the 
similar-trending faulting southward in Kentucky to the east-westward-trending 
Rough Creek Fault Zone in western Kentucky is unclear, and the relationship may 
not be a close one. 

Method of Study 

The faults in Perry and Spencer Counties were mapped by using subsurface 
information from all available petroleum, coal, mineral, and stratigraphic test holes 
on file at the Indiana Geological Survey. Both Perry and Spencer Counties have 
been explored extensively for oil and gas since the late 1920s. About 3,000 
petroleum tests have been drilled in Spencer County, and about 650 have been 
drilled in Perry County. Subsurface control, however, is sparse in those areas 
where the faults are mapped. Most of the holes were drilled with cable tools, and 
geophysical logs, which are particularly useful in well-to-well stratigraphic correla- 
tion, are not common. Driller's logs provide the basic data in southern Spencer 
County, where there are no visible surface expressions of the faults. In Perry 
County, however, where the upper Chesterian rocks are exposed in the rugged 
topography of the Crawford Upland, several good marker beds crop out and pro- 
vide the basis for detailed surface mapping. 

Correlations of electric logs and driller's logs generally uncover only those 
faults with more than about 20 feet of vertical displacement. Displacements of 
faults mapped on the surface were calculated by the difference in elevation of close- 
ly spaced outcrops of marker beds rather than by direct mapping of the fault 
planes, which are mostly covered by unconsolidated materials. 

Some outcrops mapped near faults in Perry County showed steeply inclined 
bedding, some dipping in opposite directions than would be expected from normal 
drag features near the faulting. This suggests that the faulting is complex, that 
more than one fault plane is in a zone, and that beds dip eratically between the 
planes. We also found that inclined beds of sedimentary structures could be 
mistaken easily for fault-affected beds, especially at the Mississippian- 
Pennsylvanian unconformity. Some previous investigators may have been misled 
by such structures. 

Fault Descriptions 

Faults in Perry and Spencer Counties, Indiana, trend N. 5° E to essentially an 
east-west direction with north-northeast being the most prevalent trend (Figure 1). 
The faults are all normal type, with high-angle dips on the fault planes. The Little 
Hurricane Island Fault was the only fault where a dip calculation was possible, and 
it was greater than 70°. In general, the faults in Spencer County are downthrown 
eastward, and the faults in Perry County are downthrown westward. 

Our depictions of the faults as single planes are oversimplications in that 
there is undoubtedly more complex faulting near major faults. This is borne out in 
the area near Cannelton where several small faults were observed in at least one of 
the early coal mines near the Hawesville Fault. Also, a small accessory fault 



Geology and Geography 




Figure 1. Map showing faults in southern Spencer and Perry Counties, Indiana. 



creates a 1,000 foot wide graben associated with the major Little Hurricane Island 
Fault (Figure 2). 

Isopachous maps do not indicate any thickening or thinning of stratigraphic 
units due to differential sedimentation on opposite sides of faults; therefore, there 
is no indication of growth faulting. The following described Little Hurricane Island, 
Africa, and German Ridge Faults are newly named in Indiana. The Hawesville and 



A Northwest 



Little Hurricane 
Island Fault 



Southeast A 



Africa Fault 



500 



600 



ff 700- 



800 



900 



1100- 



1200- 




Figure 2. Cross section showing faulting in southern Spencer County, Indiana, 
See Figure 1 for line of section. 



326 Indiana Academy of Science 

Indian Creek Faults are extensions of faults mapped in Kentucky; their names 
have been adopted for formal use in Indiana. 

Hawesville Fault 

The Hawesville Fault enters Indiana in sec. 16, T. 7 S., R. 3 W., from Hancock 
County, Kentucky, and trends N 5° E for 272 miles. The fault was named for the 
small river town of Hawesville, Kentucky (9). The upthrown side is to the east, and 
the maximum vertical displacement is 75 feet in sec. 16, T. 7 S., R. 3 W. The throw 
as expressed on the top of the Cypress Formation (Mississippian) decreases north- 
ward to 25 feet in less than 2 miles (Figure 3). The fault is not mapped north of sec. 
33, T. 6 S., R. 3 W., where regional dips of 40 feet per mile are on the top of the 
Cypress Formation. 

Indian Creek Fault 

The Indian Creek Fault enters Indiana in sec. 12, T. 7 S., R. 3 W., from Han- 
cock County, Kentucky, and trends N 27° E up the Deer Creek Valley about 472 
miles. The upthrown side of the fault is to the east. The fault was named for Indian 
Creek, an Ohio River tributary in Hancock County, Kentucky (9). Maximum ver- 
tical displacement in Indiana is 90 feet in sec. 12, T. 7 S., R. 3 W., on outcrops of the 



500 



400 



300 



200 



100 



B West 

Hawesville 
Fault 



East B 



German Ridge 
Fault 



Sea Level 



100 



200 



300 




Figure 3. Cross section showing faulting in southern Perry County, Indiana. See 
Figure 1 for line of section. 



Geology and Geography 327 

base of the Negli Creek Limestone Member of the Tobinsport Formation 
(Mississippian) and in the subsurface on the top of the Cypress Formation. Throws 
decrease to 65 feet on top of the Cypress Formation in sec. 6, T. 7 S., R. 2 W. 
(Figure 3), and gradually decrease to less than 20 feet in sec. 21, T. 6 S., R. 2 W. A 
drag feature exists on the west side of the road in the SE1/4NW1/4NE1/4 sec. 6, T. 
7 S., R. 2 W., in the Mount Pleasant Sandstone Member of the Tobinsport Forma- 
tion. 

German Ridge Fault 

The German Ridge Fault trends N 30° E about 8 miles, and the upthrown side 
of the fault is to the east. The fault was named after the highland area known as 
German Ridge and is not shown on mapping by the U.S. Geological Survey in Ken- 
tucky. The southern end of the fault extends to the Ohio River in sec. 8, T. 7 S., R. 2 
W., and trends north-northeastward 8 miles before dying out in sec. 1, T. 6 S., R. 2 
W. (Figure 1). The maximum vertical displacement is 60 feet in the NW1/4 sec. 23, 
T. 6 S., R. 2 W., on the top of the Cypress Formation and the Vienna Limestone 
Member of the Branchville Formation (Mississippian). The throw decreases to the 
southwest from sec. 23, T. 6 S., R. 2 W., to 30 feet on top of the Cypress Formation 
in sec. 33, T. 6 S., R. 2 W. (Figure 3). 

Little Hurricane Island Fault 

The Little Hurricane Island Fault in Spencer County, Indiana, trends N 35° E 
from sec. 11, T. 8 S., R. 7 W., about 5 miles to sec. 20, T. 7 S., R. 6 W. (Figure 1). The 
upthrown side is to the west. The fault was named for Little Hurricane Island in 
the Ohio Rvier. 

The major fault has a short parallel fault about half a mile long in the NW1/4 
sec. 1, T. 8 S., R. 7 W., and SW1/4 sec. 36, T. 7 S., R. 7 W. The splinter fault is up- 
thrown to the east and has 30 feet of displacement on top of the Cypress Formation 
(Figure 2). The Johnson Kelly No. 1 Howard well in the NW1/4NW1/4 sec. 1, T. 8 
S., R. 7 W., has about 20 feet of missing section in the Sample Formation (Mississip- 
pian) due to faulting at a depth of about 1,465 feet. The dip of the fault plane is 
greater than 70°. The major fault has a maximum vertical displacement of 50 feet 
on the top of the Cypress Formation in sec. 1, T. 8 S., R. 7 W. (Figure 2), and throw 
decreases northeastward and southwestward. 

Africa Fault 

The fault was named for the hamlet of Africa in sec. 10, T. 8 S., R. 6 W. The 
Africa Fault trends N 40° E from sec. 25, T. 8 S., R. 7 W., on the Ohio River to the 
NE1/4 sec. 9, T. 8 S., R. 6 W., where the trend changes to eastwest, leaving Spencer 
County, Indiana, in sec. 11, T. 8 S., R. 6 W., and entering Daviess County, Kentucky 
(Figure 1). The E. M. D. & G. Drlg. No. 1 Snyder well in NE1/4NE1/4NE1/4 sec. 19, 
T. 8 S., R. 6 W., has about 40 feet of section faulted out in the Sample Formation at 
a depth of 1,470 feet. The maximum vertical displacement based on interpretation 
of structure on top of the Cypress Formation is 75 feet in sec. 19, T. 8 S., R. 6 W. 

Figure 2 shows decreasing throw on the fault with depth. The vertical 
displacement is about 60 feet on top the Vienna Limestone Member and 35 feet 
on top of the Renault Formation (Mississippian). The throw decreases 25 feet in 460 
feet of vertical section, which suggests that the fault is dying out at depth. 

Deeper Faulting 

In the faulted areas of Perry and Spencer Counties, only one test hole has 



328 Indiana Academy of Science 

penetrated below upper Valmyrean (Mississippian) rocks. The lack of sufficient 
deep subsurface data, therefore, does not provide a good basis for determining 
deep faulting in lower Paleozoic strata or basement rocks, and the relatively small 
displacement of rocks faulted in the Chesterian section compared with that of rocks 
faulted in the Wabash Valley Fault System does not enhance the possibility of 
deeper faulting. Rudman and others (12) have postulated a possible basement scarp 
underlying the Wabash Valley Fault System, and more recently Braille and others 
(4) have suggested a basement rift feature in southwestern Indiana. Involvement of 
the faults in Perry and Spencer Counties as an expression of the eastern boundary 
of any such model does not seem justified. 

Literature Cited 

1. Ashley, G. H. 1899. The coal deposits of Indiana: Indiana Dept. Geology and 
Nat. Resources Ann. Rept. 23, p. 1255-1298. 

2. Ault, C. H., D. M. Sullivan and G. F. Tanner. 1980. Faulting in Posey and Gib- 
son Counties, Indiana: Indiana Acad. Sci. Proc. for 1979, v. 89, p. 275-289. 

3. Bergendahl, M. H. 1965. Geology of the Cloverport Quadrangle, Kentucky- 
Indiana and the Kentucky part of the Cannelton Quadrangle: U.S. Geol. 
Survey Geol. Quad. Map GQ-273. 

4. Braille, L. W. and others 1978. An intergrated geophysical and geological 
study of the tectonic framework of the 38th Parallel Lineament in the vicinity 
of its intersection with the extension of the New Madrid Fault Zone: West 
Lafayette, Ind., Purdue Univ., 67 p. 

5. Cox, E. T. 1872. Perry County: Indiana Geol. Survey Ann. Repts. 3 and 4, p. 
61-143. 

6. Goudarzi, G. E. and A. E. Smith. 1971. Geological map of part of the 
Owensboro West Quadrangle in Daviess County, Kentucky: U.S. Geol. Survey 
Geol. Quad. Map GQ-890. 

7. Gray, H. H., W. J. Wayne and C. E. Wier. 1970. Geologic map of the 1° x 2° 
Vincennes Quadrangle and parts of adjoining quadrangles, Indiana and Il- 
linois, showing bedrock and unconsolidated sediments: Indiana Geol. Survey 
Regional Geol. Map 3. 

8. Hughes, John H. 1951. The geology of the Deer Creek Fault area, Perry Coun- 
ty, Indiana [unpub. M.A. thesis]: Bloomington, Indiana Univ., 31 p. 

9. Johnson, W. D., Jr. and A. E. Smith. 1968. Geological map of part of the 
Owensboro East Quadrangle in Daviess County, Kentucky: U.S. Geol. Survey 
Geol. Quad. Map GQ-751. 

10. Mayfield, F. M. and D. B. Chisholm. 1952. Geologic and structure map of 
Hancock County, Kentucky: Kentucky Geol. Survey, Ser. IX, Map. 

11. Owen, Richard. 1861. Report of a geological reconnaissance of Indiana, made 
during the years 1859 and 1860, under the direction of the late David Dale 
Owen, M. D., State Geologist: Indianapolis, Ind., H. H. Dodd and Co., 368 p. 

12. Rudman, A. J. and others 1956. Geology of basement in Midwestern United 
States: Am. Assoc. Petroleum Geologists Bull., v. 49, p. 894-904. 

13. Sullivan, D. M., C. H. Ault and J. N. Stellavato. 19_. The Wabash Valley 
Fault System in Indiana and its economic implications: Kentucky Geol. Survey 
Ser. X, Spec. Pub. . 

14. Tanner, G. F., J. N. Stellavato, and J. C. Mackey 1980. Map of South- 
western Gibson County, Indiana, showing structure of Cypress Formation 
(Mississippian); Indiana Geol. Survey Misc. Map 29. 

15. Tanner, G. F., J. N. Stellavato and J. C. Mackey. 1980. Map of Northern 
Posey County, Indiana, showing structure of Cypress Formation (Mississip- 
pian): Indiana Geol. Survey Misc. Map 30. 

16. Tanner, G. F., J. N. Stellavato and J. C. Mackey. 1980. Map of Southern 
Posey County, Indiana showing structure of Cypress Formation (Mississip- 
pian): Indiana Geol. Survey Misc. Map. 31. 



HISTORY OF SCIENCE 

Chairman: Everett F. Morris 
Dean, Louisiana State University, Alexandria, Louisiana 71304 

Chairman-Elect: Gene Kritsky 
Department of Biology, Tri-State University, Angola, Indiana 46703 

ABSTRACT 

Charles Lyell's Geologic Observations in Indiana, 1846. R. William Orr, Depart- 
ment of Geography and Geology, Ball State University, Muncie, Indiana 

47306. Sir Charles Lyell visited Indiana in the Spring of 1846 during his second 

trip to North America. He traversed the southern edge of the state along the Ohio 
River first stopping at Mount Vernon from where he traveled overland to New 
Harmony. Here, accompanied by David Dale Owen, Lyell examined exposures of 
Pennsylvanian rocks and studied in situ stumps of Sigillaria nearby at Blairsville. 
He continued his journey from Evansville and traveled to New Albany where he 
examined the classic exposures of Devonian coralline limestone at the Falls of the 
Ohio. Lyell's concepts and understanding of the origin of coal, nature and distribu- 
tion of coal swamp vegetation, intercontinental correlation by means of fossils, con- 
struction of ancient "reefs," and the nature of loess deposits all were influenced by 
his studies of Indiana geology. 



329 



Mendel and the Origin of Species 

Gene Kritsky 

Department of Biology 

Tri-State University 

Angola, Indiana 46703 

Gregor Johann Mendel, the monk who is credited for unlocking the secrets of 
genetics, is an enigmatic figure in the history of biology. Moreover, Mendel's in- 
terest in Darwin and evolution, while well documented, is open for reinterpreta- 
tion. 

Most of the information about Mendel comes from his biographer Hugo litis 
(6), who first read about Mendel while a schoolboy in Briinn. (litis' interest is likely 
local pride; Mendel performed his experiments at a monastary outside Briinn.) litis' 
biography contains many questionable extrapolations. Mendel's interest in Darwin 
also was explored by Gavin de Beer (3). De Beer was the first to study Mendel's 
margin notes in the Origin of Species by Charles Darwin, but he referred to litis 
for some basic information regarding Mendel's interest in Darwin and evolution. 
The most active worker of Mendel history today is Vitezslav Orel of the Mendel 
Museum at the monastary in Briinn. Orel's work on Mendel's interest in evolution 
is critical of de Beers' comments concerning Mendel's not declaring his support of 
Darwinian evolution (9). 

Thus, the purpose of this paper is to examine Mendel's interest in evolution 
and Darwin, and to determine the influence Darwin may have had on Mendel's 
work. 

Important to the scenario is when and why Mendel started his research on 
garden peas. litis (6) has refuted the idea that Mendel, being a priest, was opposed 
to evolution and started his research to disprove it. The chronology of Mendel's 
research shows that Mendel started his work in 1856, three years before Darwin's 
Origin of Species was published. Mendel in a letter to Carl Nageli stated his ex- 
periments took place during 1856 to 1863. Therefore, Darwin could not have been 
the initial reason for Mendel's research. 

A clue to why Mendel started his research may be found upon reading 
Mendel's paper, Experiments on Plant Hybridization (7). In Mendel's paper one 
finds the names Gartner and Kolreuter often being cited. Gartner wrote a com- 
prehensive monograph in 1849 entitled Versucke and Beobachtungen ilber die 
Bastarderzeugung im Pflanzenreich. His experimental design was similar to 
Mendel's in that he performed hundreds of crosses but observed the effects of 
crosses not on one or two characteristics but several characters in many species (5). 
Mendel cited Gartner eighteen times in his paper (7). Kolreuter was a forerunner of 
Gartner, having published his major work on hybridization in 1761. Kolreuter 
studied the hybridization of different varieties of tobacco. Mendel cited Kolreuter 
six times (7). It is clear that what interested Mendel was the action of hybridization 
and how it related to species formation. Not only did Mendel often cite Gartner and 
Kolreuter but also Darwin cited the two botantists several times in the Origin of 
Species. 

The period from 1863 to 1865 when Mendel had completed his experiments, 
reviewed his data, and read his paper, appears to be the time when Darwin had his 
greatest influence on Mendel's work. It was at this time that Mendel obtained his 



History of Science 331 

copy of the Origin of Species. Mendel owned the second German translation 
published in 1863 under the name Uber die Entstehung der Arten im Thier und 
Pflanzenreich (2). The second German edition was based on the third English edi- 
tion published in 1861 (1). The fact Mendel read the text is established by the 33 
pages that Mendel marked (9). It was Mendel's habit to write page numbers down 
in the back cover and to mark some pages. A list of the pages marked by Mendel 
was published by de Beer (3). 

A review of Mendel's marginalia will reveal an interest in several points. 
First, Mendel cited three pages where Darwin defined creation and evolution. On 
page one of the Origin of Species' Historical Sketch, Darwin wrote "the great ma- 
jority of naturalists believe that species are immutable productions, and have been 
separately created. . . .few naturalists, on the other hand, believe that species 
undergo modification, and that existing forms have descended by true generation 
from preexisting forms." On page 6, still in the Historical Sketch, Darwin said that 
"by the word 'creation' the zoologist means a process he knows not what." Mendel 
also marked page 16, where he read, "the view which most naturalists entertain. . . . 
that every species have been independently created is erroneous." It is perfectly 
logical that a priest would be interested in such comments. It is possible he discuss- 
ed evolution with Klacel, another priest at the monastary who was interested in 
natural philosophy (6). 

Mendel also noted pages that included comments relating to his experimental 
design. On page 76, Darwin calculated the doubling time of some plants and 
animals. While Mendel was not necessarily concerned with rates of increase, he 
was interested in statistics and the application of mathematics to biological prob- 
lems. 

Mendel was also concerned with Darwin's comments about hybridization 
research. Page 43, one of the page numbers Mendel wrote on the back cover of the 
book, includes a discussion about the skill of gardeners. Darwin commented, "I 
have seen great surprize expressed in horticultural works at the wonderful skill of 
gardeners, in having produced such splendid results from such poor materials." 
Mendel also noted page 111, where Darwin examined the flower and pollination of 
the pea family. 

Many of the pages that Mendel marked concerned Darwin's view about 
varieties and how they related to species. Darwin's view was that "species are only 
strongly marked and permanent varieties." This theme was discussed in detail in 
several pages Mendel marked. On page 63, Darwin asserted, "Certainly no clear 
line of demarcation has as yet been drawn between species and subspecies. . . .be- 
tween subspecies and well marked varieties or between lesser varieties and in- 
dividual differences. These differences blend into each other in an insensible 
series." Moreover, Darwin stated that the individual differences are only slightly 
effected by climate and food. On page 57, recorded by Mendel in the back cover, 
Darwin wrote that "individual differences, are highly important for us, for they are 
often inherited." Mendel also noted page 63, where Darwin carried this argument 
further: "I look at individual differences ... as being the first step toward such 
slight varieties . . . And I look at varieties which are in any degree more distinct 
and permanent, as steps leading to more strongly marked and permanent varieties; 
and these latter, as leading to subspecies and to species." Mendel could appreciate 
the relevance of his work on hybrids and how variation would be involved in Dar- 
win's view on the origin of species. What Mendel did was to quantify how this 
variation moved from generation to generation. 



332 Indiana Academy of Science 

Likely the greatest surprize in the pages Mendel marked are the passages 
that indicate Darwin knew, in general terms, Mendel's conclusions. Mendel 
discovered the specifics about heredity because he worked with single characters. 
Darwin and Gartner worked on groups of characters. Therefore, it was more dif- 
ficult to spot the specific ratios Mendel described. 

Mendel marked pages 287 and 303, where Darwin correctly noted that some 
characteristics of the parent are dominant in the first generation. Darwin wrote 
that some individuals "have a remarkable power of impressing their likeness on 
their hybrid offspring." Clearly Darwin had noted that some traits were dominant, 
but his concern was in hybrids between different species and not individual genes. 
Darwin also knew that characters reappeared from generation to generation. 
Mendel likely read with interest page 302, because he marked the passage where 
Darwin dealt with the difference between the first and second generation from a 
hybrid cross. Darwin wrote, "The slight degree of variability in hybrids from the 
first cross or in the first generation, in contrast with their extreme variability in 
the succeeding generations is a curious fact and deserves attention." Darwin had 
put his finger on one of Mendel's important contributions. Namely, individuals in 
the first generation of a cross are more similar to each other because of 
heterozygous genes. When individuals from the first generation are crossed a se- 
cond time, one gets more variability. Mendel demonstrated that fact in a quan- 
titative manner, but Darwin already knew about increased variability in subse- 
quent generations in a qualitative sense five years earlier. 

How did Mendel view these comments in the Origin? It is my opinion that the 
previous works depicting Mendel as a critical evaluator of evolution are wrong. I do 
not see Mendel as a forceful individual. We must recall that Mendel failed his high 
school examination. We also must take with a grain of salt litis' comments that 
Mendel was a richer person for this experience (6). Mendel may have been a good 
teacher, but such a rebuke would not likely increase one's confidence as a scientist 
or researcher. 

It is easy to envision Mendel as the active scientist because we are looking at 
Mendel's actions as he read the Origin of Species and wrote his paper. But this 
description of Mendel should not be extrapolated into thinking of him as the domi- 
nant individual. I see Mendel as reading the Origin of Species and finding his basic 
conclusions already in Darwin's work. With the Origin of Species in mind, Mendel 
wrote his paper incorporating Darwin's philosophy. This action of trying to make 
one's research relevent with current trends in biology is still common today. 

Evidence for this thesis occurs throughout Mendel's paper (7). In the introduc- 
tory remarks Mendel states that his procedure of studying hybridization is "the 
only right way by which we can finally reach the solution of a question, the impor- 
tance of which, cannot be overestimated in connection with the history of the evolu- 
tion of organic forms." Later Mendel states, "For the history of the evolution of 
plants this circumstance is of special importance, since constant hybrids acquire 
the status of new species." Clearly, Mendel was placing his work within the 
mainstream of evolution. 

Moreover, Darwin's influence on Mendel is also seen in Mendel's discussion 
about varieties and species. Mendel wrote, "It has so far been found to be just as 
impossible to draw a sharp line between the hybrids of species and varieties 
themselves as between species and varieties themselves." This is very similar to 
what Darwin wrote on page 63, "Certainly no clear line of demarcation has as yet 
been drawn between species and subspecies . . . and varieties." 






History of Science 333 

The contention that Mendel was trying to fit his work into evolution and not 
trying to answer problems of heredity is supported in a paper by Robert Olby (8). 
Olby discusses the likelihood that Mendel did not realize he had discovered the 
basic principles of genetics but was only concerned with how hybrids related to 
species formation. This, as Olby points out, allows one to understand why no one 
recognized the genetic importance of Mendel's work. Since Mendel's basic conclu- 
sions about dominant and recessive traits, Fj constancy and F 2 variability were 
already known; therefore, Mendel discovered nothing new. Obly maintains that 
Mendel's work became important when 20th century biologists read 35 years of ad- 
vances into it. 

litis (6) claimed in his biography of Mendel that Mendel agreed with natural 
selection, but felt something was missing. litis even quoted a close friend of Mendel 
who said the name Darwin never crossed Mendel's lips. litis further quoted a col- 
league of Mendel who claimed Mendel said, "My time will come." Doubt has to be 
thrown on these recollections made years after Mendel's death. The former com- 
ment about never mentioning Darwin is in error because in his letters to Nageli, 
Mendel mentioned Darwin by name (9). The latter quotation, "My time will come," 
could have been made after Mendel read Darwin's book Variation in Animals and 
Plants under Domestication. Mendel received that text at least three years after 
he wrote his paper on hybridization. The book, Variation in Animals and Plants 
under Domestication, contains five notes by Mendel but is mostly uncut (9). In 
Variation, Darwin took a wrong turn with regard to how traits moved from genera- 
tion to generation. 

The fact that Mendel was an evolutionist is further substantiated in his letters 
to Nageli. In 1877, Mendel wrote, "If such be the real state of affairs, spontaneous 
hybridization in Hieracium must be ascribed to temporary disturbances which if 
frequently repeated or persistent must even lead to the disappearance of the 
species in question, whereas one of another more favourably organized hybrid off- 
spring better adapted to extant telluric and cosmic conditions might succeed in 
maintaining itself in the struggle for existence and might thus persist for long 
periods of time, until at length overtaken by the same fate" (6). In another letter 
Mendel wrote, "When several are competing i.e. pollen grains, we can probably 
assume that only the strongest ones succeed in effecting fertilization" (9). 



Summary 

It is established from Mendel's copy of the Origin of Species, his paper Ex- 
periments on Plant Hybridization, and his letters to Nageli that Mendel was not 
opposed to evolution, but a supporter of it. The timetable of events indicated that 
Mendel started his research in 1856 and continued it until 1863. During 1863-1865, 
while he was summarizing his research, Mendel received his copy of the Origin of 
Species. The pages he marked indicate his interest in varieties as they relate to 
species and also contain in qualitative terms his conclusions about dominant versus 
recessive traits and what happens in the Fj and F 2 generations. 

With this verification of his work by Darwin, Mendel wrote his paper placing 
it into the mainstream of evolution as it was studied in the 1860s. Mendel was 
possibly not the "priest who held the key to evolution" as Loren Eiseley (4) claimed, 
not an individual who was skeptical about evolution as litis claimed (6), but rather 
was converted to evolution and wrote his paper as a supporter of Darwinism, try- 
ing to demonstrate the importance of his research to the current trends in biology. 



334 Indiana Academy of Science 

Acknowledgements 

I thank Everett Wilkie of the Lilly Library at Indiana University and Frank 
Gyorgyey of the Yale University Medical School Library for their help in obtaining 
the second German edition of the Origin of Species. I also thank Peter Hippensteel, 
Simon Black, James Bourke, and Jack Nortrup for their many comments. 

Literature Cited 

1. Darwin, C. 1861. On the Origin of Species by means of Natural Selection, or 
the preservation of favoured races in the struggle for life. John Murray, Lon- 
don. 

2. Darwin, C. 1863. Uber die Entstehung der Arten im Thier-und Pflanzen-Reich 
durch naturliche Ziichtung: 2te Auflage, Stuttgart. 

3. de Beer, G. 1966. Mendel, Darwin, and Fisher (1865-1965). Notes and Records 
of the Royal Soc. London 19 (2): 192-226. 

4. Eiseley, L. 1958. Darwin's Century. Doubleday and Co., New York. 

5. Gartner, C. F. 1849. Versuche and Beobachtungen uber die Bastarderzeu- 
gung im Pflanzenreich. Stuttgart. 

6. Iltis, H. 1966. Life of Mendel. Hafner Publ. Co., New York. 

7. Mendel, G. 1963. Experiments in Plant Hybridization. Harvard University 
Press, Cambridge. 

8. Olby, R. 1979. Mendel no mendelian? Hist. Sci. 17:53-72. 

9. Orel, V. 1971. Mendel and the evolution idea. Folia Mendeliana 6:161-172. 



Arthur J. Phinney, M.D., 
Indiana's First Subsurface Geologist 

Walter H. Pierce 

Department of Geography/Geology 

Ball State University 

Muncie, Indiana 47306 

In the words of John B. Patton, State Geologist of Indiana, Arthur J. Phin- 
ney's paper entitled "The Natural Gas Field of Indiana" was "one of the earliest 
and best in the annals of petroleum geology" (1). Though Phinney lived to be 92 
years old, only 15 years were spent doing geologic research, and during these 
years he maintained his medical practice in Muncie, Indiana. Edgar Wesley Owen 
observed: "Many highly trained geologists were less productive than the amateur 
Phinney." (2) 

Arthur John Phinney was born in Russell Township of Geauga County, Ohio 
in late August of 1850. He was raised on a farm 20 miles east of Cleveland (3). He 
received his early education in high school and the Geauga Seminary, an academy 
of that area. After attending the academy, he entered Oberlin College. After com- 
pleting two terms (one year) at Oberlin College he went on to Allegheny College in 
Meadville, Pennsylvania for two more years, where he reached the junior year. 
During these college studies he taught in the common schools at Geauga County 
and surrounding area for six years. He concentrated on the sciences and 
mathematics during these early years of education (4). 

In 1875 at the age of 25 he began his medical studies. He attended Wooster 
University (4) and Western Reserve Medical School (3). From one of these institu- 
tions he left to enter Pulte Medical School in Cincinnati, Ohio. He graduated from 
Pulte Medical School in 1877 (3, 4). 

Phinney began a short medical practice in Galion, Ohio and then, in October 
1878, he went to Muncie, Indiana where he began a long medical practice (4). In 
Muncie he joined the local Literary and Scientific Association of Muncie. Between 
1878 and 1882 he gave two talks to that society, one entitled "Evolution from a 
Geologic Standpoint" and another entitled the "Earth and Its Motions (5)." These 
talks indicate that Dr. Phinney had developed a certain amount of expertise in 
geology prior to beginning his geologic studies in east-central Indiana. 

There is no indication when Dr. Phinney first became interested in geology; 
however, a short biography states that he had been a student of geology for many 
years. His first recognition as a geologist was in 1881 when he was employed to 
make a geological survey of Delaware County (4). At this time John Collett was 
state geologist. Apparently the geologic studies were begun in association with 
Phinney's botanical studies of the area. For his study of Delaware County Phinney 
received $30.00 (6). 

Dr. Phinney continued his medical practice while doing geology. He completed 
five separate articles for the Indiana Geological Survey. The first article appeared 
in the 11th Annual Report in 1881 and was entitled the "Geology of Delaware Coun- 
ty" (7). In the following year a report on the "Geology of Randolph County" (8) and a 
"Catalogue of the Flora of East Central Indiana" (9) appeared in the 12th Annual 
Report. Apparently Phinney was paid $100.00 for the report on Randolph County. 
For the 13th Annual Report Phinney completed a general report on Henry and por- 
tions of Randolph, Wayne and Delaware Counties (11). 

335 



336 Indiana Academy of Science 

In these regional reports Dr. Phinney correctly identified the age of the 
bedrock as Silurian. He recognized the glacial origin of the drift and identified the 
Muncie esker, for which he used the Scandinavian spelling, assar. He recognized 
that many river valleys were underfit and additionally that many of the original 
prairies in this region represented the position of ancient lakes now filled with 
peat. To help prevent summer peat fires, he suggested improved methods of peat 
extraction. In the initial phases of subsurface work related to the gas field, he 
recognized bedrock valleys buried beneath the glacial till. 

Concurrently with his regional work the Gas Boom began. Gas was discovered 
in Ohio in November 1884. A bore, originally drilled for coal at Eaton in Delaware 
County was deepened, and as a result gas was discovered in Indiana in September 
1886. 

In Maurice Thompson's 1886 report on Natural Gas in Indiana he quoted Dr. 
A. J. Phinney of Muncie who in his words had ". . . proved himself a very able and 
painstaking assistant,. . ." The following words show Dr. Phinney's attention to 
detail when describing the early Eaton Well: 

"The gas has some odor, though not very unpleasant, it burns without 
smoke, and is thought to be free of sulfur. The roar produced by the 
escaping gas can, under favorable conditions, be heard at a distance of 
two miles. A two-inch pipe was extended from an elbow at the top of the 
casing to a point 18 feet above the derrick, or 90 feet from the ground. 
Another two-inch pipe was extended horizontally from the elbow about 
60 feet from the well. Both pipes were furnished with a T, giving four 
places for escape of the gas. When lighted the flame from each was about 
10 feet long. The light could easily be seen from Muncie, twelve miles 
south, and I was told that it had been seen twenty miles" (12). 

At this time Dr. Phinney was 36 and must have been the acknowledged expert 
on the geology of the region. In the words of W. J. McGee: 

"When the drill began its operations in Indiana it fell to Dr. Phinney, an 
amateur geologist of Muncie, to observe and study the boring records; 
and to him belongs the credit for commencing and carrying well forward, 
at his own cost a comprehensive investigation of the data brought to light 
by the test borings in that State (13)." 

After the first seven paying wells had been drilled in Indiana, Phinney 
published a short article entitled "Facts About Gas" in the January 28, 1887 In- 
dianapolis News. Phinney obtained records for seventeen wells. He determined 
the elevation of the "mouth of each bore" and from the well record found the eleva- 
tion of the Trenton. From this data he determined that a structural arch existed in 
the area of the gas field and that the arch had an axis which ran southeast-to- 
northwest. He also surmised that outcrops of Niagaran limestone might be used as 
a clue to the location of the arch. By analogy to other gas fields which were control- 
led by "anticlinals or archs", he implied that the arch controlled the occurrence of 
gas in the Trenton (14). 

By July 1887, Dr. Phinney had constructed a preliminary map of the Indiana 
Gas Field. This map was published in a weekly periodical entitled the American 
Manufacturer and Iron World on September 2, 1887. This remarkable map must 
qualify as the first structural contour map of the Trenton Oil Field in Indiana (15). 
This map precedes Orton's 1888 Ohio Geological Survey publication "Oil and Gas in 



History of Science 337 

Ohio" (16) and Orton's 1889 publication "The Trenton Limestone as a Source of 
Petroleum and Inflamable Gas in Ohio and Indiana" which was published in the 9th 
Annual Report of the United States Geological Survey (17). 

The map is a representation of the elevation of the top of the Trenton. That 
portion of the Trenton which lies above sea level is shown in ruled lines, the portion 
from sea level to 100 feet below sea level is shown in dashed lines, and that portion 
which is lower than 100 feet below sea level is left white, without pattern. 

He termed the 100-foot below sea level line the "dead line", and suggested 
strongly that efforts to find gas outside the "dead-line" would be less fortunate. 

In October, 1887 Dr. Phinney published a more comprehensive article entitled 
"The Gas Territory of Indiana" in the periodical, Petroleum Age. In this article he 
reviewed the literature on the Cincinnati Arch or Anticlinal and began to refine 
further his concept of the trap, reservoir, and pressure aspects of the gas field (18). 

These articles caught the attention of Major John Wesley Powell, then direc- 
tor of the United States Geological Survey. Powell sent W. J. McGee to visit Phin- 
ney in Muncie during the height of drilling activity in 1887. As a result of this visit, 
Phinney was appointed United States Geological Surveyor for the Indiana Gas 
Field. Arrangements were made to complete the work for publication. A portion of 
this work was published in the United States Geological Survey Mineral 
Resources for 1887 in 1888 (19). In Phinney's words: "the study took place over a 
period of three years" (4). It was complete in March of 1890 and the work resulted 
in a major contribution to the 11th Annual Report of the United States Geological 
Survey published in 1891 (20). 

The American Association of Petroleum Geologists Memoir 5 (The 
Sourcebook of Petroleum Geology) refers to this work nine times (21). The source- 
book attributes to Orton and Phinney the first use of the word "trap" in petroleum 
geology. Phinney may well have been the first to use it in the form of a noun. Dr. 
Phinney was one of the first geologists to state clearly that hydrostatic pressure 
played a role in gas migration. He also recognized that the gas in the Trenton Field 
accumulated from broader areas adjacent to the Cincinnati Arch. Thus Phinney 
recognized the concept which would later be called "gathering area" and that gas 
could migrate considerable distances. 

One of the most striking contributions of Dr. Phinney was his list of requisites 
for a gas field. A modern list of this kind has been published by Momper for oil and 
gas (22). 

1. Effective Source 

2. Carrier System 

3. Reservoir 

4. Effective Seal 

5. Trap 

6. Timing 

7. Preservation 

Phinney's list includes the following: 

1. A carbonaceous stratum within which the hydrocarbons are generated 

2. A porous rock overlying it or adjacent to it 

3. An impervious cover to prevent its escape 

4. The presence of the necessary structural relief to enable a separation of 
the gas, oil, and salt water to take place 



338 Indiana Academy of Science 

5. The absence of extensive deformation and fracture of the strata that 
would afford avenues of escape 

6. Absence of outcrop whereby the products might escape 

Compared to Momper's 1978 list, the only thing missing from Phinney's 1891 list is 
the carrier system. 

Also included in this volume (20) were nicely printed color maps of the geology 
of the state, with the Trenton Gas Field superposed over the geology; and a struc- 
tural contour map of the entire state, called a hypsographic map. This map may 
represent the first structural contour map of an entire state. 

At the age of 41, Dr. Phinney rightfully could consider himself a physician, an 
accomplished geologist, and a botanist. He considered music his only pastime (4). 
Perhaps it was in order to simplify his life that he determined to devote himself en- 
tirely to his medical career. It seems this was his decision since in the fall of 1892 he 
sold his collection of fossils, minerals, and land shells which numbered greater than 
17,000 species, "all classified and labeled (4)." 

He continued to practice medicine and probably moved to Indianapolis in 1907 
or 1908 where he practiced medicine for a few years before retiring. Phinney 
outlived his wife, Mary, who died in 1923 and is buried in Beech Grove Cemetery of 
Muncie. 

He was a charter member of the Indiana Academy of Science and spoke at the 
50th anniversary meeting of the Academy in 1934 (3). He retained an interest in 
non-medical science, in the evolution of religious beliefs, in the origin of the 
American Indian, whom he believed to be a cross between the black and brown 
man, and in the problems of irrigation in the western deserts. Phinney was 
reported to have written 1,200 pages of manuscript which he hoped to have 
published (3). The last few years of his life were spent in the Masonic Home in 
Franklin, Indiana (4). During this time his only daughter Louise passed on in 1940. 
Dr. Phinney died on May 26, 1942. He was buried in Muncie's Beech Grove 
Cemetery on May 28, 1942 along side his wife and daughter in the Little Family 
plot. At the present, neither his daughter's nor his grave has markers. 

It is unfortunate that McGee employed the term amateur to describe Dr. Phin- 
ney. This adjective has too many shades of meaning. Not only was Dr. Phinney an 
expert in the geology of east-central Indiana, but his understanding of the entrap- 
ment of hydrocarbons was probably further advanced than that of most geologists 
of his time. His description of the Trenton Gas Field possibly represents the 
earliest description of a carbonate stratigraphic trap (20). One can only imagine 
what contribution Dr. Phinney might have made to geology had he devoted a larger 
part of his life to this science. 

Though his geologic career was short Dr. Arthur J. Phinney became Indiana's 
first subsurface geologist. 

Literature Cited 

1. Patton, J. B. 1978. Notable contributions of early Indiana geologists. Pro- 
ceedings of Indiana Academy of Science. 88:278-281. 

2. Owen, E. W. 1975. Trek of the oil finders: a history of exploration for 
petroleum. Amer. Assoc, of Petroleum Geologists, Tulsa, Oklahoma. Memoir 
6. 1647 p. 

3. Edington, W. E. 1942. Necrology. Proceedings of the Indiana Academy of 
Sciences. 52:7-8. 



History of Science 339 

4. Bowen, A. W. 1894. A portrait and biographical record of Delaware County, 
Indiana. A. W. Bowen & Co., Chicago. 807 p. 

5. Helm, T. B. 1881. History of Delaware County. Kingman Brothers, Chicago. 
303 p. 

6. Collett. J. 1882. Financial statement of receipts and expenditures. 12th An- 
nual Report of Indiana Department of Geology and Natural History. 11-16. 

7. Phinney. A. J. 1881. Geology of Delaware County. 11th Annual Report of In- 
diana Department of Geology and Natural History. 126-149. 

8. Phinney. A. J. 1882. Catalogue of the flora of central-eastern Indiana. 12th An- 
nual Report of Indiana Department of Geology and Natural History. 196-236. 

9. Phinney. A. J. 1882. Geology of Randolph County. 12th Annual Report of In- 
diana Department of Geology and Natural History. 177-195. 

10. Phinney. A. J. 1883. Geology of Grant County. 13th Annual Report of Indiana 
Department of Geology and Natural History. 138-153. 

11. Phinney. A. J. 1886. Henry County and portions of Randolph, Wayne and 
Delaware. 15th Annual Report of the Indiana Department of Geology and 
Natural History. 97-116. 

12. Phinney, A. J. 1886. The Eaton gas well in Natural Gas by Maurice Thomp- 
son. 15th Annual Report of the Indiana Department of Geology and Natural 
History. 321-322. 

13. McGee, W. J. 1891. Letter of Transmittal. U. S. Geological Survey Annual 
Report No. 11, pt. 1. 587-588. 

14. Phinney. A. J. 1887. Facts about gas. The Indianapolis News. January 28th. 

15. Phinney. A. J. 1887. The Indiana gas field. American Manufacturer and Iron 
World. September 2nd. 15-19. 

16. Orton, E. 1888. The Trenton Limestone as a source of oil and gas in Ohio. Ohio 
Geological Survey Report No. 6. 101-310. 

17. Orton, E. 1889. The Trenton Limestone as a source of petroleum and in- 
flamable gas in Ohio and Indiana. U. S. Geological Survey Annual Report No. 
8. 475-662. 

18. Phinney. A. J. 1887. The gas territory of Indiana. The Petroleum Age. 
6:1743-1747. 

19. Phinney, A. J. 1888. Natural Gas in Indiana. U. S. Geological Survey Mineral 
Resources for 1887. 485-489. 

20. Phinney, A. J. 1891. The natural gas field of Indiana. U. S. Geological Survey 
11th Annual Report, pt. 1. 579-742. 

21. Dott, R. H., and M. J. Reynolds. 1969. Sourcebook for Petroleum Geology. 
Amer. Assoc, of Petroleum Geologists Memoir 5, Tulsa, Oklahoma. 471 p. 

22. Momper. J. A. 1978. Oil migration limitations suggested by geological and 
geochemical considerations in Physical and chemical constraints on petroleum 
migration. Amer. Assoc, of Petroleum Geologists continuing education course 
notes series No. 8. B-l-B-60. 



MICROBIOLOGY AND MOLECULAR BIOLOGY 

Chairman: Donald A. Hendrickson 
Department of Biology, Ball State University, Muncie, Indiana 47306 

Chairman-Elect: Dorothy Adalis 
Department of Biology, Ball State University, Muncie, Indiana 47306 

ABSTRACTS 

A Simplified Method for Storing Anaerobic Bacteria. William W. Baldwin and 
Ming Tseng, Indiana University Medical School, Northwest Center for Medical 
Education, Gary, Indiana 46408 and Marshall Landay, St. Margaret Hospital, 

Hammond, Indiana 46320. The recommended method for storage of anaerobic 

bacteria is at room temperature in chopped meat broth lacking carbohydrates. 
There are two problems associated with the use of chopped meat broth. One is its 
complicated preparation and the other problem is its turbidity. This turbidity 
necessitates gram staining to determine if growth has occured and risks oxygen 
and microbial contamination. We have developed an agar medium in which we have 
successfully stored anaerobic bacteria for ten months. The medium is composed of 
Blood Agar Base (Difco) 40g/£, Yeast Extract (Difco) 5g/f, and cysteine HCf 0.05 gll 
The medium is prereduced and anaerobically sterilized in 13 x 100mm screw cap 
tubes and inoculated with a loop long enough to reach the bottom of the tube. The 
tube is capped and sealed with parafilm and incubated at 37° C for 2 days, then 
stored at room temperature. 

The Effects of a Hexaflora on the Morphology of the Gerbil. Kenneth F. Bartizal, 
Margaret H. Beaver and Bernard S. Wostmann, Lobund Laboratory, University 

of Notre Dame, Notre Dame, Indiana 46556. The use of the gerbil as an animal 

model in biomedical research has increased dramatically. The gerbil is of interest 
because its serum cholesterol is quite responsive to the level of dietary cholesterol 
yet, when maintained in a state of hypercholesterolemia, no evidence of plaque for- 
mation is observed. The gerbil is also an excellent model to study bile acid (BA) 
metabolism because of the similarity of its BA pattern to that of man. 

The effects of an intestinal microflora on cholesterol and BA metabolism are 
substantial. In order to separate microbial factors from physiological mechanisms 
that influence sterol metabolism, germfree (GF) and gnotobiotic (GN) gerbils were 
obtained. 

GF gerbils were caesarian derived; but because of cecal enlargement, they 
failed to reproduce. A pair of GF gerbils were associated with a defined six 
member murine derived microflora that was known to have very limited effects on 
the primary BA pattern of rats. This hexaflora, consisting of Lactobacillus brevis, 
Streptococcus faecalis, Staphylococcus epidermidis, Bacteroides fragilis, var. 
vulgatus, Enterobacter aerogenes, and a Fusobacterium sp. allowed the GN ger- 
bils to reproduce and a colony of hexaflora associated hexa-gerbils has been 
established. In order to broaden the colonies genetic base, caesarian sections were 
performed to introduce additional gerbils when a female hexa-gerbil was nursing a 
new litter. 

The hexa-gerbil has a much higher serum cholesterol level than the CV gerbil 
when fed a (0.1%) cholesterol supplemented diet. The serum lipoprotein pattern in- 
dicates that in GN gerbils harboring a microflora with very limited secondary BA 

340 



Microbiology and Molecular Biology 341 

modifying capability, the animal accumulates cholesterol in the more suspect 
VLDL and LDL fractions, while the HDL fraction stays the same. 

Morphological data on 4 to 7 month old male hexa-gerbils indicate a sizable 
reduction of the cecum when compared to its GF counterpart. However, both small 
and large intestine weighed substantially more than in the CV gerbil. Also, the size 
of the thymus was significantly reduced, suggesting a certain amount of stress im- 
parted by this murine-derived microflora. 

This chronic inflammation of the mucosa is most likely related to the presence 
of Enterobacter aerogenes, Staphylococcus aureus or the Fusobacterium sp. pre- 
sent in this limited component hexaflora. As monoassociates these organisms have 
been known to induce inflammation of the intestinal mucosa. 

At present, studies of tissue histology and quantitative bacteriology of the 
gerbil's gut are underway to determine the possible localization of the hexaflora 
components and their ensuing effects on the intestinal mucosa. 

Isolation, Identification, and Reinfection of Cephalosporium spp. and Two Bacteria 
Associated with Wilt in Helianthus annuus. Larry Herrman and Mary Lee 
Richeson, Department of Biological Sciences, Indiana University-Purdue Univer- 
sity at Fort Wayne, Fort Wayne, Indiana 46805. Recent discovery of wilt in the 

agronomically important crop sunflower has resulted in economic loss. In the fall of 
1979 the authors observed a severely lodged and diseased field of the oil producing 
sunflower near Auburn, Indiana. A fungus and two bacteria were isolated from 
lesions on the stem of diseased plants. The fungus was identified by microscopic com- 
parisons as Cephalosporium spp. The isolate was grown on Sabourard and potato 
dextrose agar at various temperatures. Optimum growth on agar is 20-25° C. 

Cephalosporium is a known plant pathogen causing wilt in several woody 
plants. As far as the authors know, it has not been reported before as a pathogen of 
sunflower. The two bacteria identified as Erwinia carotovora and Xanthomonas 
campestris are also known plant pathogens. Two different times of inoculation and 
four methods of inoculation have established that the sunflower is not susceptible 
to the disease produced by Cephalosporium in the seedling stage. Ten day old 
seedlings were scratched, sprayed, and injected with a suspension of 
Cephalosporium. None of the treatments indicated any symptom of disease. In ad- 
dition to the actual inoculation of the organism into the seedlings, the soil in which 
some of the sunflowers were to be grown was treated with a suspension of 
Cephalosporium. Seeds planted in this treated soil also produced healthy plants 
with no sign of disease. Plants inoculated with the Cephalosporium suspension at 
the onset of senescence, however, showed rapid hyphae growth and rapid spread of 
the fungus over the plant. Inoculation method did not seem to be a factor in 
establishment of the disease. The soil treatment was not used this time. Other 
etiological investigations with the two bacteria isolates and the Cephalosporium 
isolate are under way. 

Dissolved Oxygen Profile of an Aerobic Bio-Reactor. Robert H. L. Howe. West 
Lafayette, Indiana 47906. The dissolved oxygen profile of an aerobic bio- 
reactor is discussed. Emphasis with respect to the different utilization and oxida- 
tion rates of different substrate components is explained. A mathematical model 
for the prediction of the dissolved oxygen concentration progression is introduced. 

Search for Phosphoproteins in Bacillus subtilis. Mark 0. Oster and Sylvia Brehm 
Oster. Indiana State University, Terre Haute, Indiana 47809. Several proteins 



342 Indiana Academy of Science 

in many biological systems are post translationally modified by phosphorylation 
resulting in altered activity. In order to investigate the possibility that some pro- 
teins in Bacillus subtilis may be phosphorylated, we grew B. subtilis in the 
presence of radioactive P 32 orthophosphate (5mC/l). Crude extracts were obtained 
by treatment with lysozyme, incubated with DNase and RNase and subjected to 
chromatography on Sephadex G-50. The excluded fraction, containing chiefly pro- 
teins, was analyzed for the incorporation of P 32 by scintillation counting. The nature 
of the phosphate attachment was investigated. Neither P 32 -labelled phosphoserine 
nor phosphothreonine could be detected in the protein fraction after hydrolysis in 
6N HC1 at 110 C for 4 hr and chromatography on Dowex-1 with authentic stan- 
dards. The P 32 -labelled material is stable at 37° C for 30 min over a wide pH range 
from 2 to 12 but labile at pHl and pH13 and above. Under the same conditions, the 
P 32 -labelled product is stable in 1M neutral hydroxylamine indicating that the at- 
tachment is not an acyl phosphate, phosphohistidine nor phosphyolysine. The 
possibility of other covalent linkages to protein and polyphosphates is still under 
investigation. 

Rate Studies on Microbial Chitin Decomposition in the Freshwater Habitat. Toni 

L. Poole and Carl E. Warnes, Department of Biology, Ball State University, Mun- 

cie, Indiana 47306. Microbial action on particulate chitin was analyzed in an 

East-central Indiana borrow pit lake. Experiments were conducted to determine 
the effects of water depth, particle size, mesh size of nylon bag containing chitin, 
and season on the decomposition process. The majority of chitinolytic bacteria 
isolated were classified as actinomycetes and pseudomonads. Results indicate that 
the sediment-water interface is the most active site of chitin mineralization in the 
lake environment. The samples seeded during summer showed the fastest rate of 
decomposition with greater than 50% weight loss after two weeks and greater 
than 95% weight loss after 7 weeks incubation in situ. Samples seeded during the 
spring showed considerably slower activity with only a 25% weight loss after 9 
weeks incubation in situ. Larger particle size appears to slow the mineralization 
process. 

Isolation and Identification of Two Bacteria Associated with Wilt in Helianthus an- 
nuus. Mary Lee Richeson and Larry G. Herrman, Department of Biological 
Sciences, Indiana University-Purdue University at Fort Wayne, Fort Wayne, In- 
diana 46805. Recent discovery of wilt in the oil-producing sunflower, Helian- 
thus annuus, has resulted in agricultural losses. Two types of bacteria and a fungus 
were isolated from lesions on the stem of diseased plants. In this study, the two 
bacterial isolates were identified. Microscope studies of morphology revealed both 
isolates to be small gram-negative asporogenous motile rods. Capsules were pre- 
sent. Temperature optima were 20-25 C. One isolate produced large, white, domed, 
mucoid colonies on nutrient agar containing 5% sucrose. The other isolate produced 
yellow pigmented colonies. These and other observations together with data from 
approximately twenty biochemical tests indicate that one organism is probably Er- 
winia carotovora, the other, Xanthomonas campestris. Both are known plant 
pathogens. Xanthomonas has not been associated with disease in sunflower. Er- 
winia are facultative anaerobes of the family Enterobacteriaceae which produce 
acid and gas from glucose but are otherwise fermentatively erratic. Growth at 36 C 
and lack of yellow pigment establish this as a member of the Carotovora group. 
Xanthomonas are strict aerobes, never fermentative. Ecologically, they are the 
plant pathogen components of the Pseudomonadaceae. Preliminary etiological ex- 
periments indicate no definite symptoms appear if sunflower plants are inoculated 






Microbiology and Molecular Biology 343 

with Erwinia as seedlings. Other etiological investigations with Erwinia, Xan- 
thomonas, and the fungal isolate are under way. 

Methodology for Measuring Nitrogen Fixation by Acetylene Reduction in Beijerin- 
ckia and Klebsiella in Stream Litter Decomposition. Dilipkumar Vyas, Depart- 
ment of Biology and Bruce Storhoff, Department of Chemistry, Ball State Univer- 
sity, Muncie, Indiana 47306. The nitrogen-fixing abilities of Beijerinckia and 

Klebsiella have been determined by gas chromatographic measurements of 
acetylene to ethylene ratios in incubated cultures. Commercially available 
acetylene was purified and added to the cultures, and the amount of ethylene pro- 
duced was measured using a Varian 1400 Chromatograph equipped with a flame 
ionization detector and a 1 m x 2.5 mm I.D. column packed with phenylisocyanate 
on Porasil. Calibration standards consisting of mixtures of acetylene, ethylene and 
argon were prepared using vacuum line techniques. The chromatographic and data 
treatment procedures will be presented in detail. 

Investigation of the Role of Beijerinckia and Klebsiella as Nitrogen Fixers in 
Stream Litter Decomposition. Dilipkumar Vyas, Donald Hendrickson, and Carl 
Warnes. Department of Biology and Bruce Storhoff, Department of Chemistry, 

Ball State University, Muncie, Indiana 47306. The role of Beijerinckia spp and 

Klebsiella spp in stream litter decomposition was investigated using acetylene 
reduction method. Beijerinckia and Klebsiella were isolated during winter and 
spring of 1980, from leaves of sugar maple (Acer saccharum), placed in Bell Creek, 
Muncie, Indiana. Nitrogen free media was used to isolate Klebsiella. Both of these 
organisms were streaked separately on nitrogen free agar slant tubes, flushed with 
argon and acetylene added. Gas chromatographic analyses were conducted on the 
inoculated tubes after incubation for 24 hours at 32° C. Beijerinckia spp. isolated 
from January, February and April fixed 4.48 x 10 2 moles and Klebsiella spp. 
isolated during the same time period was between 1.47 to 8.10 x 10 8 moles. This 
study indicates that Beijerinckia is a better N 2 fixer than Klebsiella in stream lit- 
ter decomposition. 



Bacterial Community Dynamics and Species Diversity of Slime Producing 
Bacteria at Selected Sites on the Ohio River 

Donald A. Hendrickson 
Department of Biology, Ball State University, Muncie, Indiana 47306 

Introduction 

This study deals with the seasonal population trends of bacteria in the Ohio 
River intake water at Cardinal Plant, Clifty Creek Station, Kyger Creek Station 
and Tanner's Creek generating facilities from May, 1978 through April, 1979. A 
weekly comparison was made between slime forming total aerobic bacteria and 
slime forming aerobic bacteria and between gram negative aerobic bacteria and 
slime forming gram negative aerobic bacteria. 

Study Sites 

The study sites chosen for this study were all located on the Ohio River in the 
southern part of Ohio and Indiana. These sites were selected because they were all 
locations of electric generating facilities that had previously experienced slime 
buildup in their condenser tubes; Cardinal Plant located near Brilliant, Ohio; Kyger 
Creek Station near Gallipolis, Ohio; Tanner's Creek Plant near Lawrenceburg, In- 
diana; and Clifty Creek Station near Madison, Indiana. 

Methods and Materials 

A. Sample Collection for Bacteriological Analysis 

Triplicate samples of intake water were collected weekly in sterile Nalgene 
bottles. These samples were packed in blue ice and shipped in 2 inch 
styrofoam packing by UPS to our laboratory. 

B. Bacteriological Analysis: 

Bacteriological analyses of the samples were prepared by making serial dilu- 
tions of all the samples with phosphate buffered water. One tenth ml of the 
serial dilutions was plated on Standard Methods Agar and MacConkeys Agar. 
The plates were incubated at ambient temperature (22°C) for five days. 

Results 

Seasonal Bacterial Populations 

Bacterial population numbers in intake water varied seasonally from May, 
1978 through April, 1979 (Figures 1-8). The total bacterial population fluctuated 
widely from about 10,000/ml to near or in excess of 100,000/ml. 

Total slime bacterial population numbers in intake water at all sites were 
generally much lower than total bacterial population numbers except from late 
July through early November, 1978 when they were quite similar (Figures 1, 3, 5, 
7). Total slime bacteria at all stations was usually less than 20% of total bacterial 
population except during the July to November period when they made up about 
50 to 95% of the population. 

Total gram negative bacteria were usually about a factor of 10 lower than 
total bacteria with few exceptions (Figures 2, 4, 6, 8). Population levels never ex- 
ceeded 100,00/ml and they commonly fluctuated above and below 10,000/ml. 

Total gram negative bacterial population levels in intake water at sites were 
similar to total gram negative bacterial levels from June through September 

344 






Microbiology and Molecular Biology 



345 




■ * * 




TOTAL SLIME 



MAY JUN JUL AUG SEP OCT NOV DEC JAN FEB MAR APR 

Figure 1. Bacterial population dynamics (total and total slime) at Cardinal Plant, 
May, 1978 through April, 1979. 



T r 




MAY JUN JUL AUG SBP OCT NOV DEC JAN FEB MAR APR 

Figure 2. Bacterial population dynamics (total gram negative and slime gram 
negative) at Cardinal Plant, May, 1978 through April 1979. 



346 



Indiana Academy of Science 




TOTAL SLIME 



MAY JUN JUL AUG SEP OCT NOV DEC JAN FEB MAR APR 

Figure 3. Bacterial population dynamics (total and total slime) at Clifty Creek 
Station, May, 1978 through April, 1979. 



1 1 1 1 1 1 1 1 1 r 




MAY JUN JUL AUG SEP OCT NOV DEC JAN FEB MAR APR 

Figure 4. Bacterial population dynamics (total gram negative and slime gram 
negative) at Clifty Creek Station, May, 1978 through April, 1979. 



Microbiology and Molecular Biology 



347 




s s TOTAL 



TOTAL SLIf" 



J L 



MAY JUN JUL AUG SEP OCT NOV DEC JAN FEB MAR APR 

Figure 5. Bacterial population dynamics (total and total slime) at Kyger Creek 
Station, May, 1978 through April 1979. 



i 1 1 r 




A TOTAL GRAM NEGATIVE 



SLIME GRAM NEGATIVE 



MAV JUN Jin, AUG SEP OCT NOV DEC 



JAN FEB MAR APR 



FIGURE 6. Bacterial population dynamics (total gram negative and slime gram 
negative) at Kyger Creek Station, May, 1978 through April 1979. 



348 



Indiana Academy of Science 




' a-4 



TOTAL SLIME 



MAY JUN JUL AUG SEP OCT NOV DEC JAN FEB MAR APR 

Figure 7. Bacterial population dynamics (total and total slime) at Tanner's Creek 
Plant, May, 1978 through April 1979. 




>i - * 



TOTAL GRAM NEGATIVE 



SLIME GRAM NEGATIVE 



MAY JUN JUL AUG SEP OCT NOV DEC JAN FEB MAR APR 

Figure 8. Bacterial population dynamics (total gram negative and slime gram 
negative) at Cardinal Plant, May, 1978 through April 1979. 



Microbiology and Molecular Biology 349 

(Figures 2, 4, 6, 8). The number of slime gram negative bacteria also tended to drop 
relative to total gram negative bacteria from October, 1978 through April, 1979, 
although population levels were never below 100/ml. 

Species Dominance 

In addition to isolating and counting total aerobic bacteria, total aerobic slime 
producing bacteria, gram negative bacteria isolated on MacConkeys agar, and 
slime producing bacteria isolated on MacConkeys agar, all slime producing bacteria 
isolated were identified by API rapid diagnostic procedure. Frequency of occur- 
rence of bacteria isolated is shown in Table 1. The most frequently isolated slime 
producing bacterium was Pseudomonas fluorescens. Greatest species diversity oc- 
curred in the cooling water of all plants from mid July to early September. A 
change in species composition was observed during this reporting period. 
Acinetobacter calco, var antitrat, a Bacillus species and various Pseudomonads 
were not present at the beginning of the reporting period but were dominant later. 
The genus Enterobacter occurred in all plants during the summer reporting 
period. In addition to the seasonal variation in species composition at each report- 
ing site, there were also variations in species composition between study sites for 
the same reporting date. 

Discussion and Conclusions 

Bacteriological data and river water temperature were collected from May 1, 
1978 to April 30, 1979 at four electric generating facilities on the Ohio River: Car- 
dinal Plant, Clifty Creek Station, Kyger Creek Station, and Tanner's Creek Plant. 
Major conclusions are as follows: 

1. Total bacterial numbers were variable between about 10,000/ml and 
100,000/ml during the sample period. 

2. Total slime bacteria dominated bacterial populations from mid July through 
September. 



Table 1. Frequency of occurrence of slime forming bacteria isolated in the 
cooling waters of Cardinal Plant, Clifty Creek Station, Kyger Creek Station and 

Tanner's Creek Plant. 





CARDINAL 


CLIFTY 


KYGER 


TANNER'S 


Acinetobacter calco 










var. anitrat. 


80 a 


90 


83 


67 


Aeromonas spp. 


43 


31 


25 


39 


Alcaligenes spp. 


95 


73 


87 


98 


Bacillus spp. 


28 


27 


38 


24 


Enterobacter agglomerans 


8 


6 


2 


31 


Enterobacter spp. 


10 


21 


12 


25 


Flavobacterium breve 


1 


1 


1 


6 


F lav o bacterium spp. 


10 


21 


12 


25 


Moraxella spp. 


3 


4 


4 


6 


Pasteurella multocida 


10 


21 





20 


Pseudomonas aeruginosa 


15 


4 


12 


29 


Pseudomonas fluorescens 


100 


75 


94 


82 


Pseudomonas multophilia 


5 


25 


4 


6 


Pseudomonas paucimobilis 


5 


15 


8 


6 


Pseudomonas spp. 


95 


98 


92 


67 


Xanthamonas spp. 


3 


33 








a = percent of samples containing 


bacterium 









350 



Indiana Academy of Science 



The greatest bacterial species diversity occurred in the cooling water of all 

plants from mid July to early September. 

A change in slime bacterial species composition of the cooling water occurred 

at all reporting stations during the reporting period. Variations in species 

composition between the sample sites for the same reporting date were also 

observed. 



Acknowledgements 

This work was funded by American Electric Power Service Corporation, Ohio 
Valley Electric and Indiana-Kentucky Electric Corporation. 



Analysis of Factors Influencing Population Dynamics of Slime Forming Bacteria at 
Selected Sites on the Ohio River 

Donald A. Hendrickson and Thomas S. McComish 

Department of Biology, Ball State University 

Muncie, Indiana 47306 



Introduction 

Biofouling of condenser tubes at Clifty Creek Station, an electric power 
generating facility on the Ohio River in Southern Indiana, occurred in March 1976. 
Analysis of condenser tubes revealed biofouling was due to bacterial adhesion in- 
side condenser cooling tubes. Bacteria isolated from condenser tubes produced 
copious layers of slime. Similar biofouling due to bacteria occurred at Cardinal 
Plant and Tanner's Creek Plant on the Ohio River in Southern Ohio and Indiana. 

This study was undertaken to evaluate seasonal population trends of bacteria 
in the Ohio River intake water at Cardinal Plant, Clifty Creek Station, Kyger 
Creek Station and Tanner's Creek generating facilities from May, 1978 through 
April, 1979. A weekly comparison was made between total aerobic bacteria and 
slime forming aerobic bacteria and between total gram negative aerobic bacteria 
and slime forming gram negative aerobic bacteria. Statistical models, involving 
selected measured variables at sites, were used to gain insight to factors influenc- 
ing bacterial populations in intake water. 

These results represent the first attempt at elucidating factors influencing 
bacterial populations in Ohio River intake water at the sites noted. This paper in- 
cludes summarized information from a more comprehensive evaluation of aquatic 
bacterial populations by McComish and Hendrickson (2). Funding for this work was 
provided by American Electric Power Service Corporation, Ohio Valley Electric, 
and Indiana-Kentucky Electric Corporation. 

Study Sites 

The study sites were located on the Ohio River in Southern Ohio and Indiana. 
These sites were selected because they were all locations of electric generating 
facilities previously experiencing bacteria slime buildup in condenser tubes; Car- 
dinal Plant near Brilliant, Ohio; Kyger Creek Station near Gallipolis, Ohio; 
Tanner's Creek Plant near Lawrenceburg, Indiana; and Clifty Creek Station near 
Madison, Indiana. 

Methods and Materials 

Triplicate samples of intake water were collected weekly, kept cold, and usual- 
ly processed within 48 hours. Bacteriological samples were prepared by plating 
water samples on Standard Methods Agar and MacConkeys Agar. Plates were in- 
cubated at ambient temperature (72°F, 22C) for 5 days and total bacterial and slime 
bacterial counts obtained. Slime formation was confirmed by stringy consistency of 
the colony and capsule formation. Each bacterium confirmed as a slime producer 
was identified using API 20E strips. Chemical analyses were performed according 
to Standard Methods for the Examination of Water and Wastewater (1). Lim- 
nological characteristics examined for inclusion in statistical evaluation and model 
formulation included intake water temperature, pH, biochemical oxygen demand 
(BOD ), ammonia, alkalinity, hardness, turbidity, total dissolved solids, and sulfate. 

351 



352 



Indiana Academy of Science 



Statistical analyses were conducted using a DEC-10 system and standard 
analytical computer procedures. 

Results and Discussion 

Seasonal Bacterial Populations 

Bacterial population numbers in intake water varied seasonally from May 
1978 through April 1979. The total bacterial population fluctuated widely from 
10,000/ml to near or in excess of 100,000/ml. 

Total slime bacterial population numbers in intake water at all sites were 
generally much lower than total bacterial population numbers except from late 
July through early November 1978 when they were quite similar. This strong 
trend of total slime bacteria population dominance is evident when total slime 
bacteria are evaluated as a percentage of total bacteria at sites (Figures 1-4). Total 
slime bacteria at all stations were usually less than 20% of total bacteria except 
during the July to November period noted when it made up from about 50-95%. 

Total gram negative bacteria essentially followed the same seasonal pattern 
as described for total bacteria at each site. Population levels were, however, lower 
by about a factor of 10 with only a few exceptions. 

Discriminant Analysis Models 

Discriminant analysis was applied to prediction of bacterial population percen- 
tages for total slime bacteria of total bacteria and for gram negative slime bacteria 
of total gram negative bacteria. This was accomplished by using two population 
groups (group 1: 0-49.9% and group 2: 50.0-100.0%) for percent slime bacteria 
(Table 1) and for percent gram negative slime bacteria (Table 2). 



• TOTAL SLIME 



— — -■ GRAM NEGATIVE SLIME 
A INTAKE TEMPERATURE 




Figure 1. Percent total slime bacteria and gram negative slime bacteria with 
temperature at Cardinal Plant from May, 1978 through April, 1979. 



Microbiology and Molecular Biology 



353 



• TOTAL SLIME 

■ GRAM NEGATIVE SLIME 

k INTAKE TEMPERATURE 




eo § 

g 

i 



MAY JUN JUL AUG SEP OCT NOV DEC JAN FEB MAR APR 

Figure 2. Percent total slime bacteria and gram negative slime bacteria 
temperature at Clifty Creek Station from May, 1978 through April 1979. 



with 




MAY JUN JUL AUG SBP OCT NOV DEC 



MAR APR 



Figure 3. Percent total slime bacteria and gram negative slime bacteria with 
temperature at Kyger Creek Station from May, 1978 through April, 1979. 



354 



Indiana Academy of Science 




FEB MAR 



Figure 4. Percent total slime bacteria and gram negative slime bacteria 
temperature at Tanner's Creek Plant from May, 1978 through April, 1979. 



with 



Percent slime bacteria of total bacteria were classified into group one correct- 
ly 71% of the time and into group two correctly 93% of the time for data combined 
from stations over the 12 month period using intake water temperature, ammonia, 
and BODg as discriminators (Table 1). Models for individual sites ranged from 
74-100% correct prediction of either group. Significant variables (F test, 95% level) 



TABLE 1: Models of discriminant analysis allowing classification of percent 

slime bacteria of total bacteria into two population groups 0-49* and 50.0-100.0; 

using intake water temperature, ammonia, and BOD$ at sites combined and 

separated for data collected from May, 1978 through April, 1979. 







Functions 




Predicted Correctly (%)* 


Models 


Temperature 


Ammonia 


BOD 5 


Constant 


Group 1 
0-49.9 


Group 2 
50.0-100.0 


Combined 
Overall 


Combined Sites 


-0.0624 


-0.1845 


0.0990 


4.3552 


71 


93 


77 


Cardinal Plant 


0.0561 


-2.6043 


** 


-2.9990 


74 


85 


78 


Clifty Creek 
















Station 


0.0455 


-1.7031 


0.6674 


-3.5519 


90 


100 


92 


Kyger Creek 
Station 


0.0641 


** 


0.1621 


-4.0990 


81 


92 


85 


Tanner's Creek 
Plant 


0.0175 


-2.6727 


-0.3927 


2.5690 


86 


100 


91 



* Classified using discriminant model for prediction. 

** Variable rejected for use in formulating discriminant model. 



Microbiology and Molecular Biology 



355 



Table 2. Models of discriminant analysis allowing classification of percent 

gram negative slime bacteria of total gram negative bacteria into two 

population groups. 0-49.9* and 50.0-100.0* using intake water temperature, 

ammonia, and BOD$ at sites combined and separated for data collected from 

May, 1978 through April 1979. 



Functions 



Predicted Correctly (%)* 



Models 


Temperature 


Ammonia 


BOD 5 


Constant 


Group 1 
0-49.9 


Group 2 
50.0-100.0 


Combined 
Overall 


Combined Sites 


0.0621 


•» 


** 


-3.9687 


68 


95 


74 


Cardinal Plant 


0.0644 


-2.4820 


»* 


-3.8011 


73 


93 


83 


Clifty Creek 
Station 


-0.0271 


4.5277 


»* 


0.0296 


74 


100 


76 


Kyger Creek 
Station 


0.0651 


-0.3363 


0.1746 


-3.6879 


81 


92 


84 


Tanner's Creek 
Plant 


0.0826 


-1.3090 


-0.6628 


-2.9407 


91 


100 


94 



Classified using discrimination model for prediction 
Variable rejected for use in formulating discriminant model. 



allowing group predictions included: intake water temperature, ammonia and 
BOD,- at Clifty Creek Station and Tanner's Creek Plant, but only intake water 
temperature and ammonia at Cardinal Plant, and intake water temperature at 
Kyger Creek Station. 

Results were similar for percent gram negative slime bacteria of total gram 
negative bacteria as shown for total bacteria (Table 2). Gram negative slime 
bacteria were correctly predicted for groups one and two for combined data in 68 
and 95% of the cases, respectively, using only intake water temperature as a 
discriminator. Models for individual sites ranged from 73-100% correct prediction. 
Models for Kyger Creek Station and Tanner's Creek Plant utilized intake water 
temperature, ammonia, and BODj- while Cardinal Plant and Clifty Creek Station 
used only intake water temperature and ammonia. 

It is significant to note that intake water temperature was an important 
discriminator at all stations. Inspection of graphs of percent slime bacteria and 
gram negative slime bacteria (Figures 1-4) show intake water temperatures in- 
creasing to about 80°F (27C) by mid July. After mid July and into September, in- 
take water temperatures remained high (at or near 80°F, 27C). This was the same 
period of total slime bacteria and gram negative slime bacteria population 
dominance at all sites. While other variables such as ammonia and BODr were in- 
volved in predictive models, temperature is clearly the major factor influencing the 
aquatic bacterial population. It appears that the slime forming bacteria were better 
adapted than non slime forming bacteria to the prevailing ecological conditions, 
especially temperature, during the sample period. 

Summary and Conclusions 

Selected physical, chemical and bacteriological data were collected from May 
1, 1978 to April 30, 1979 at four electric generating facilities on the Ohio River: Car- 
dinal Plant, Clifty Creek Station, Kyger Creek Station and Tanner's Creek Plant. 
The data were used in computer analyses to formulate statistical models. Seasonal 
population trends were examined in relation to selected measured variables. Major 
conclusions are as follows: 



356 Indiana Academy of Science 

1. Total bacterial numbers were variable between about 10,000-100,000/ml 
during the sample period. 

2. Discriminant models were formulated to predict slime bacteria as a 
percentage of total bacteria and to predict gram negative slime bacteria 
as a percentage of total gram negative bacteria. Water temperature was 
the major discriminating variable common to all models formulated. 
Classification of bacteria as dominant or subdominant was correctly ac- 
complished using the models from 68-100% of the time. 

3. Slime bacteria dominated bacterial populations from mid July through 
September when river intake water temperatures were above or near 
80°F (27C). Ecological conditions, especially elevated temperature, ap- 
parently favors the slime forming bacterial population of the river system 
during mid to late summer. 

Literature Cited 

1. American Public Health Association, American Water Works Associa- 
tion and Water Pollution Control Federation. 1976. Standard Methods for 
the Examination of Water and Wastewater. Amer. Pub. Health Assoc. 874. p. 

2. McComish, Thomas S. and Donald A. Hendrickson. 1979. Computer 
statistical analysis of factors influencing condenser performance at four elec- 
tric generating facilities. Final Report American Electric Power Service 
Corp., Ohio Valley Electric, and Indiana Kentucky Electric Corp. 
(mimeograph). 69 p. 



Properties of Defective Interfering Particles Induced by 
Photodynamic Treatment on Pseudorabies Virus 

I. L. Sun and D. P. Gustafson 

Department of Biological Sciencies, 

Purdue University 

West Lafayette, Indiana 47907 

Introduction 

The term "defective virus" has been applied in a number of ways to indicate 
nonreplicating particles and viruses that are structurally different and functionally 
deficient from standard virions. Van Magnus first described such influenza virions 
in 1954 as being "incomplete" (24) because they were noninfectious hemag- 
glutinating particles. That repeated passage of undiluted virus in tissue cultures 
would result in the production of defective interfering (DI) particles was 
demonstrated with herpes simplex virus (3), pseudorabies virus (1,2), and New- 
castle disease virus (NDV) (2). The alteration of DNA structure in DI particles was 
observed in HSV and PrV only. However, the interference of DI particles with the 
growth of standard virus was reported for all three viruses. It would be of con- 
siderable interest to determine the mechanism of the interference. Subsequent in- 
formation also suggested that the host cell plays a very critical role in the produc- 
tion of DI particles and the nature of their defectiveness (2,5,17,23). 

In this communication we describe isolation and characterization of a popula- 
tion of DI PrV particles prepared with a photodynamic method. 

Materials and Methods 

Pig kidney cells (PK-W2E) derived from 3-week-old pigs were used at passage 
70 in the studies. The cells were grown in flasks with Eagle's medium containing 
10% fetal bovine serum, 100 u of penicillin and 170 mg streptomycin per ml at pH 
7.4 and maintained in a similar medium containing 2% fetal bovine serum. 

The FH strain of PrV was used throughout the study. Virus infectivity was 
assayed in PK cells by determination of the 50% tissue culture infective does 
(TCID^q) or by measuring the plaque forming units per ml (pfu/ml) at 48-120 hrs. 
post exposure. 

The photodynamic treatment of virus was that described previously in detail 
by Sun et al (19). 

Infected cell cultures were frozen and thawed three times after which the col- 
lected material was centrifuged at 1500 rpm for 15 min. The virus in the super- 
natant fluids was sedimented in a sucrose gradient as described previously (19). Op- 
tical density (OD) readings were taken immediately, after drop collections, at 280 
m/i and 260 m^ with Gilford Instrument's spectrophotometer (model 2400-5). 

DNA of purified virions as described above was extracted by the sodium 
dodecyl sulfate (SDS) chloroform-isoamyl alcohol method essentially according to 
Marmur (15). 

DNA prepared from Marmur's method was centrifuged in a 5-20% sucrose 
gradient for 16 hr. at 13,000 rpm. OD reading was taken at 260 m^ Gilford Instru- 
ment's spectrophotometer (model 2400-5). 

Crossed immunoelectrophoresis was done as described by Vestergaard (22) 

357 



358 Indiana Academy of Science 

with a few modifications. Glass plates (9.4 cm by 9.4 cm) were used and were 
covered with 1.5 mm thick 1% (wt/vol) agarose (Nutritional Biochemicals Corpora- 
tion) dissolved in a buffer as described previously (20,25). The first dimensional 
electrophoresis was performed in 90 min. at 17 v/cm of gel and the second dimen- 
sional electrophoresis was completed in 15 hr. at 7 v/cm of gel. The plates were 
then dried, stained, and treated as described. (20,25). In the first-dimensional elec- 
trophoresis, the agarose gel, both a neutral detergent, Triton X-100 (0.5%) and an 
anionic detergent, sodium deoxycholate (0.05%) were incorporated. In the second 
dimensional electrophoresis, gel contained purified anti-PrV gamma-globulin 
prepared as described previously (21). 

Results 

Samples of infected cells were withdrawn at the time intervals of visible light 
irradiation indicated in Table 1. The OD of cell suspensions were then measured at 
540 mpi.. After 45 min. of irradiation, the efficiency of host-cell lysing ability dropped 
to 13% of the control which had not been treated photodynamically. Longer incuba- 
tion such as 60 min. did not make a significant difference (Table 1) in the protection 
of cells against lysis. However, the longer incubation periods have been used to 
reduce virus yields by 10 6 amounts, based on kinetic studies (19). Similarly, herpes 
simplex titers were reduced by 10 5 amounts (12). 

Table 2 showed that the addition of photodynamically inactivated, [acridine 
dye plus light (ADD] particles to a standard virus inoculum reduced the yield of in- 
fections virus by approximately 100 fold whether the cells were exposed both 
simultaneously or in tandem. Thus, it was demonstrated that ADL virions contain- 
ed a factor which interfered with the production of infections virus. The effects of 
the incubation with PrV antiserum or of UV light on the interfering ability of ADL 
particles were also tested. Exposure of ADL particles to standard antiviral serum 
for 1 hr. at 37 °C or UV light (20 watts) for 5 min. markedly reduced their interfer- 
ing ability. It is therefore clear that the interference with the synthesis of infec- 
tious virus by ADL culture fluids resides in particles which had antigenic proper- 
ties similar to that of standard PrV. 

In the experiment illustrated in Figure 1 the sedimentation profile obtained 
from sucrose gradients centrifugation (15-30%) is shown. The purified DI particles 
had a much lower sedimentation value than that of the purified standard virions. 
There was 38% reduction in protein synthesis (OD 280 m/x) and 14% reduction in 
nucleic acid synthesis (OD 260 m/*) in DI particles when compared with that of stan- 
dard virions. The ratio of OD 280 m/*/ OD 260 m^i = 1.12 in DI particles was also dif- 
ferent from that of infectious standard virions (OD 280 mlm/OD m/* = 1.33). The 
ratio also indicated that much less protein was made in the DI particles. 



Table 1. Protection of host-lysing ability by photodynamic treatment. 

Irradiation time (min.) Percent of cells being lysed 

(control) 100.0 

10 66.7 

20 30.0 

30 16.7 

45 13.3 

60 10.0 

The concentration of 0.08 uglml of dye (3,9-diamine-acridine dye) (A.D.) was used throughout the 
experiment. 



Microbiology and Molecular Biology 359 

Table 2. Capacity of photodynamically treated PrV to interfere with the 
replication of standard PrV. 

Infection Virus titer (TCID^mU 

standard PrV control (VC) 10 8 

♦ ADL 10 2 

VC + ADL (co-infection) 10 6 

VC 1 * 11 "- ADL (superinfection) 10 6 

** ADL + Antiserum (ADLS) no c.p.e. 

VC + ADLS (co-infection) 10 8 

VC 1 *""- ADLS (superinfection) 10 8 

***ADL + 5 min UV irradiation (ADLU) no c.p.e. 

VC + ADLU 10 8 

VC" 11 '- ADLU 10* 

* ADL cell cultures (6 x 10 ) were infected by 1 ml of photodynamically treated PrV (0.08 uglml of AD plus 
45 min. irradiation of visible light). 

**ADLS cell cultures were infected by 1 ml of ADL particles which had incubated for 1 hr at 37°C water 
bath with 1:20 dilution of standard PrV antiserum from swine. 
***ADLU cell cultures were exposed to 1 ml of ADL particles which had been treated with 20 watts of UV 
light for 5 min. Coinfection was performed when cultures were inoculated with both VC (m.o.i = 5) and 
ADL particles at the same time. Superinfection was performed when cultures were infected with VC first 
and ADL particles 1 hr later. The infected cultures were washed to remove unabsorbed virus as well as 
PrV antiserum. 



Effect on coinfection of pig kidney cells with ADL defective interfering par- 
ticles and standard pseudorabies virus on plaque formation was studied. Aliquots 
of each fraction collected after sucrose gradient centrifugation were then tested for 
interfering ability (pfu/ml) by comparing the progeny of cells coinfected with stan- 
dard virus and DI particles with standard virions alone as shown in Figure 2. Most 
fractions indicated about 3 log^Q reductions in the titer. Fractions 34, 35 and 36 had 
a larger measure of interference with the production of standard virions. 

Demonstrations of antigenicity in the ADL particles and quantitation of total 
antigenic proteins in them with rocket immunoelectrophoresis has been reported 
(25). However, identification of individual antigenic determinants presented in the 
immunoelectrophoretic precipitate profile, was accomplished by crossed im- 
munoelectrophoresis with the second dimensional gel containing anti-PrV swine 
sera against the purified PrV particles. Analysis of individual precipitates 
presented in standard PrV virions was illustrated previously (21). The precipitate 
pattern revealed four antigenic determinants as indicated Ag 1, Ag 2, Ag 3 and Ag 
4 which have a quantitative ratio of 19%, 10%, 20% and 51% respectively. Since 
the area outlined by each precipitate is proportional to the amount of the cor- 
responding antigenic determinant when the concentration of antibodies in the 
second-dimensional gel is kept constant, then a quantitative ratio could be deter- 
mined. 

Comparison of the precipitin profile between untreated standard virus and 
ADL particles was demonstrated in Figure 3A and Figure 3B. The untreated con- 
trol showed four antigenic precipitates (Figure 3A) as observed in our previous 
study. However, the precipition profile released from ADL particles showed only 
Ag 2 and Ag 4 to be present (Figure 3B). There were no detectable amounts of Ag 1 
and Ag 3. The defectiveness was therefore not only shown in the lack of infec- 



Indiana Academy of Science 



A 



/x 




J 1 1 1 1 I I I I I I I L 



J I I I i ' 



2 4 6 8 10 12 14 16 



2022 24 26283032 3436384042 
FRACTION NUMBER 



B 



A 



T-T-i 1 I'-f-i E 



•t ' v i 



/ \ 




2 4 68 10 12 14 16 18 2022 24 262830323436384042444 

FRACTION NUMBER 

Figure 1. Sedimentation in sucrose gradients of defective and standard virions. 
Cultures were infected with standard PrV at m.o.i. = 10 and purified in sucrose 
gradients centrifugation as described in materials and methods. Each fraction col- 
lected after centrifugation was taken O.D. readings at 260 m\i (A) and 280 mix (B). 
— : untreated control; : cultures treated with acridine dye plus visible light. 



tiveness but also in the antigenic components of ADL defective interfering par- 
ticles. 

Purified DNA was extracted by SDS — chloroform — isoamyl alcohol method 
as described by Marmur (15). A comparison of the sedimentation profile in a neutral 
sucrose gradient of purified DNA found in standard and in defective virions is 
shown in Figure 4. The study suggested that the DNA of ADL virions has a 
sedimentation value similar to that of standard virus. 

Discussion 

The formation of incomplete virus or DI particles is not an exclusive peculiarity 
of animal viruses. Similar forms have been described in DNA bacteriophage, T4 



Microbiology and Molecular Biology 361 




20 22 24 26 
FRACTION NUMBER 



Figure 2. Effect of coinfection of pig kidney cells with ADL defective interfering 
particles and standard pseudorabies virus on plaque formation. Controlled 
cultures were infected with standard PrV fm.o.i. = 5) and coinfected cultures were 
infected with standard PrV (m.o.i = 5) and ADL virions (1 ml). Cultures were 
harvested after 2-4 h. of post-infection. Results are presented as the p.fu./ml 
observed with each fraction collected from 15-30* sucrose gradients centrifuga- 

tion. — : cultures were infected with standard virus alone; ; cultures were 

coinfected with standard virus and ADL defective interfering particles. 



(13), RNA bacteriophage, Qo (16), and plant mosaic virus (14). All these DI particles 
are non-infective and have a lower sedimentation constant. However, in contrast to 
our ADL particles (Figure 4), they differ from standard virus by the absence of 
nucleic acid. 

Huang and Baltimore (9) proposed a model of DI particles in viral disease in- 
fection in which repeated cycles of infection produce a gradually increasing propor- 
tion of DI particles ultimately expressed in natural disease as persistent or latent 
infections. Since our results suggest a similar phenomenon, we have considered 
that DI particles similar in effect to the photodynamically produced ADL particles 
may be important determinants of the course of acute PrV infections and of persis- 
tent, slowly progressing pseudorabies viral disease in swine. 

Our results that ADL particles lost infectivity but retained antigenicity are 
reminiscent of an observation (24) of Von Magnus in which there was formation of 
noninfectious but hemagglutinating particles of influenza virus that were termed 
"incomplete viruses". 

According to Henle's concept (7), the formation of DI particles are assumed to 
be the result of arrested intracellular virus development as some intermediary 
stage caused by factors present either in host cells or the virus itself. In the pre- 
sent case it is possible that DI particle formation in cell cultures is the result of host 
cell enzyme systems being altered under the conditions of photodynamic treatment 
so that certain phases of intracellular virus replication are deleted. However, the 
exact mechanism by which acridine dye at very low dose (0.08 /xg/ml) prevents viral 
maturation is not well understood at present. Although, it is known that irradiation 
of protein and DNA with visible light in the presence of acridine dye leads to DNA- 
protein cross-linking (10, 18). It is also very possible that this cross-linking of viral 



Indiana Academy of Science 



4 *&- — - 








O 



Figure 3. Comparison of crossed immunoelectrophoretic patterns of 
precipitating antigens between untreated virus control and ADL treated virions. 
Immunoelectrophoresis was performed as that described in materials and 
methods. A: untreated virus control; B: ADL treated virions. 



Microbiology and Molecular Biology 



363 



0.4-1 



0.3- 



0.2- 



0.1 




oo-i — r— i — i — i — i — i — i — i — i — i — r— i — i — i — i 

2 4 6 8 12 14 16 18 20 22 24 26 28 30 

FRACTION NUMBER 

Figure 4. Sedimentation in a sucrose gradient of the purified DNA presented in 
standard and in defective virions. The virions were isolated and purified from 
cultures, as described in materials and methods. The DNA from these virions 
were extracted, as described by Marmur (13). Centrifugation was performed in a 
5-20* neutral sucrose gradient for 16 hr. at 13,000 r.p.m. — ; DNA of purified stan- 
dard PrV; : DNA of purified ADL virions. 



DNA-protein complexes may interfere with the viral maturation process and thus 
leads to biological inactivation. This may be due to factors associated with the 
viruses themselves rather than to factors associated with the host cells. 

We have applied both a neutral detergent, Triton X-100 (0.5%) and an anionic 
detergent, sodium deoxycholate (0.05%) in our two-dimensional (crossed) im- 
munoelectrophoresis. The electrophoretic mobility of amphilic proteins in standard 
virus of DI particles can be altered by so-called "charge-shift electrophoresis" (6) 
due to anodic migration and resolution of antigens are greatly improved in our two- 
dimensional immunoelectrophoresis by the incorporation of these detergents. This 
type of combination of "charged-shift" electrophoresis with regular two- 
dimensional immunoelectrophoresis thus permitted us to have better identification 



364 Indiana Academy of Science 

and comparison of antigenic determinants between standard virions and ADL par- 
ticles. 

In summary, we can say that ADL-produced DI particles differ from the fully 
active standard virions by the apparent lack of inefectivity (25), by their capacity to 
interfere with the propagation of infective virus (Table 2 and Figure 2), by having 
a lower sedimentation constant (Figure 2) and by the absence of two out of four 
antigenic determinants (Figure 3). 

Literature Cited 

1. Ben-porat, T., J. M. Demarchi and A. S. Kaplan. 1974. Characterization of 
defective interfering viral particles present in a population of pseudorabies 
virions. Virology. 60: 29-37. 

2. Ben-porat, T., and A. S. Kaplan. 1976. A comparison of two populations of 
defective, interfering pseudorabies virus particles. Virology. 72: 471-479. 

3. Bronson. D. L., G. R. Dreesman, N. Bixwal and M. Benyesh-Melnick. 1973. 
Defective virions of herpes simplex viruses. Intervirology. 1: 141-153. 

4. Cramer. W. A., and R. B. Uretz. 1966. Acridine orange sensitized photoinac- 
tivation of the capacity of Escherichia coli for bacteriophage T 4 . Virology. 28: 
142-149. 

5. Dogherty. R. M., H. S. Distefano and F. K. Roth. 1967. Virus particles and 
viral antigens in chicken tissues free of infectious avian Leukosis virus. Proc. 
Nat'l. Acad. Sci. U.S. 58: 808-817. 

6. Helenius, A. and K. Simmons. 1977. Change shift electrophoresis: simple 
method for distinguishing between amphilic and hydrophilic proteins in 
detergent solution. Proc. Nat'l. Acad. Sci. U.S. 7k: 529-532. 

7. Henle, W. and G. Henle. 1949. Studies on host-virus interactions in the chick 
embryo - influenza virus system. J. Exptl. Med. 90: 23-37. 

8. Huang. A. S., J. W. Greenawalt and R. R. Wagner. 1966. Defective T par- 
ticles of vesicular stomatitis virus. I. Preparation, morphology and some 
biologic properties. Virology. 30: 161-172. 

9. Huang. A. S. and D. Baltimore. 1970. Defective viral particles and viral 
disease processes. Nature. 226: 325-327. 

10. Kadish, L. J., D. B. Fisher and A. B. Pardee. 1967. Photodynamic inactivation 
of free and vegetative bacteriophage T 4 . Biochem. and Biophys. Acta. 128: 

57-65. 

11. Kahn, N. C. and R. K. Poddar. 1974. Photodynamic inactivation of antigenic 
determinants of single stranded DNA bacteriophage X174. J. Virol. 13: 
997-1000. 

12. Khan, N. C, J. L. Melnick, and N. Biswal. 1977. Photodynamic treatment ot 
herpes simplex virus during its replicative cycle. J. Virol. 21: 16-23. 

13. MAAL0E, 0. and N. Symonds. 1953. Radioactive sulfur tracer studies on the 
reproduction of T 4 bacteriophage. J. Bacteriology. 65: 177-182. 

14. Markham, R. 1953. "The nature of virus multiplication" p. 85. University 
Press, Cambridge. 

15. Marmur, J. 1961. A procedure for the isolation of deoxyribonucleic acid from 
micro-organisms. J. Mol. Biol. 3: 208-218. 

16. Mills, D. R., R. L. Peterson and S. Speigelman. 1967. An extracellular darwi- 
nian experiment with a self-duplicating nucleic acid molecule. Proc. U.S. Nat. 
Acad. Sci. 58: 217-224. . 

17. Roman, J. M. and E. H. Simon. 1976. Defective interfering particles in 
monolayer-propagated newcastle disease virus. Virology. 69: 298-303. 



Microbiology and Molecular Biology 365 

18. Spikes, J. D. and R. Livingston. 1969. The molecular biology of photodynamic 
actions: sensitized photoautoxidation in biological systems. Adv. Radiat. Biol. 
3: 29-121. 

19. Sun. I. L., D. P. Gustafson and G. Scherba. 1978. Comparison of pseudorabies 
virus inactivated by bromo-ethylene-imine, 60 Co irradiation and acridine dye 
in immune assay systems. J. Clin. Microbiol. 8: 604-611. 

20. Sun. I. L., J. C. Wang and D. P. Gustafson. 1979. Autigenicity of solubilized 
protein from cells infected with pseudorabies virus. Proc. Ind. Acad. Sci. 88: 
110-119. 

21. Sun. I. L. and D. P. Gustafson. 1980. Production of incomplete pseudorabies 
viruses in enucleated pig kidney cells. Proc. Ind. Acad. Sci. 89: 120-127. 

22. Vestergaard. B. F. 1973. Crossed immunoelectrophoretic characterization of 
herpesvirus hominis type 1 and 2 antigens. Acta Pathol. Microbiol. Scand. 
Sect. B. 81: 808-810. 

23. Vogt. P. K. 1967. A virus released by "Nonproducing" Rous Sarcoma cells. 
Proc. Nat'l. Acad. Sci. U. S. 58: 801-808. 

24. Von Magnus. P. 1954. Incomplete forms of influenza virus. Advan. Virus Res. 
2: 50-79. 

25. Wallis. C. and J. L. Melnick. 1964. Irreversible photosensitization of viruses. 
Virology. 23: 520-527. 



PHYSICS AND ASTRONOMY 

Chairman: Gerald P. Thomas 

Department of Physics and Astronomy, Ball State University, 

Muncie, Indiana 47306 

Chairman-Elect: L. Dwight Farringer 
1407 East Orchard Drive, North Manchester, Indiana 46962 

ABSTRACTS 

A Vaiveless Drain-Down Solar Collector System. L. Dwight Farringer, Depart- 
ment of Physics, Manchester College, North Manchester, Indiana 46962. To 

avoid cold-weather nightime freezing of water-filled solar heat collectors in a 
domestic hot water system, it is common to use either a closed-loop heat exchanger 
system with antifreeze liquid in the collector loop or an open-loop system with 
temperature controlled valves to isolate the collectors from the domestic water 
system and drain them when freezing conditions prevail. An alternative form of 
open-loop system has been devised, in which the collectors automatically drain 
whenever the circulation pump stops. From tests of a small model, it appears that 
this system may have cost, efficiency, and reliability advantages. Some 
unanswered questions will be discussed, and plans for a full-scale test system will 
be described. 

Design and Construction of a Dye Laser. J. W. Kaellner and D. R. Ober, Depart- 
ment of Physics and Astronomy, Ball State University, Muncie, Indiana 47306. 

An organic dye laser has been designed and is currently under construction. The 
dye laser consists of a mirror-lens combination and a jet stream of dye. The jet 
stream is excited transversely by a 5-watt argon laser. The open laser cavity design 
makes the device particularly useful as an instructional and demonstration tool. 
Preliminary studies will be discussed in which Rhodamine 6G is the lasing medium. 

Some Problems Involved in Obtaining Accurate Oscillator Strengths for Use in 
Astrophysics. Ronald E. Pitts, Department of Physics and Astronomy, Ball State 

University, Muncie, Indiana 47306. During the last decade, there have been 

great improvements in computers, in astronomical telescopes, and in telescope in- 
strumentation. As a result, more realistic theoretical stellar models have become 
possible, and interest has been generated in the fields of stellar evolution, 
cosmogony, and cosmology. With these new investigations has come the need for 
more accurate abundance determinations of the elements present in both the sun 
and in other stars. 

Abundance determinations are based on atomic oscillator strengths or 
f-values. Traditionally there have been difficulties in all methods of obtaining ex- 
perimentally determined f-values. These difficulties range from procuring data 
free from systematic errors to being unable to apply an accurate technique to a par- 
ticular line or lines for which f-values are needed. The introduction of lasers into 
atomic spectroscopy offers some new hope for circumventing some of these dif- 
ficulties. The f-values obtained with these new laser techniques should be of suffi- 
cient accuracy to assist in new improved abundance determinations. 

IUE Observations of the Peculiar Object BL Lacertae. Roger L. Scott, Depart 
ment of Physics and Astronomy, Ball State University, Muncie, Indiana 47306, 

366 



Physics and Astronomy 367 

Karen R. Hackney and Richard L. Hackney, Department of Physics and Astrono- 
my, Western Kentucky University, Bowling Green, Kentucky 42101. Recent 

observations of the ultraviolet spectral energy distribution of the extragalactic ob- 
ject BL Lacertae have been obtained using the International Ultraviolet Explorer 
(IUE) satellite. Low resolution (6A) IUE spectrograph^ images were obtained with 
the LWR camera (3200-1900 A) and the SWP camera (1900-1300 A) on 1979 August 
20 and 21 respectively. The discussion will compare the ultraviolet data with 
ground based optical-wavelength observations made at an earlier epoch. The obser- 
vations were made at Goddard Space Flight Center with the assistance of the IUE 
Observatory staff, and supported by NASA under grant NSG 5238. 

Active Solar Heating Systems for Residential Application in Indiana: A Com- 
parative Optimization Study. R. Taylor and R. Cosby. Department of Physics and 

Astronomy, Ball State University, Muncie, Indiana 47306. The FCHART 

method was used to size and study the performance of residential solar space and 
water heating systems for four Indiana locations. Local manufacturers or 
distributors supplied necessary data for the flat-plate collector systems selected 
for study. Applied to a new "average" Indiana house, the effects of catastrophic 
fuel increases and tax credits on system design and economics were investigated. 

A Tube-Type Photographic Sensitometer. Robert L. Tonkel and Roger L. Scott, 
Department of Physics and Astronomy, Ball State University, Muncie, Indiana 

47306. The authors have constructed and tested a "tube-type" sensitometer for 

determining the relative light sensitivity of various photographic emulsions. The 
instrument is designed after one in use at the University of Florida, and the pro- 
totype model built by D. W. Latham of the Smithsonian Astrophysical Observatory 
(Latham 1969). The discussion will include descriptions of the design and function 
of the sensitometer and its use in a program for developing and testing various 
"hypersensitization" techniques which are used to increase the sensitivity of 
photographic emulsions. 

Reference: Latham, D. W., "A Harvard-Smithsonian Tube Sensitometer," 
American Astronomical Society Photo-Bulletin, No. 1, p. 3, 1969. 



"Contra Luz Opal:" Structure and Optical Properties 

William W. Davis 
4124 North Pennsylvania Street, Indianapolis, Indiana 46205 

Introduction 

"Contra luz" means in Spanish "against the light." This opal is so named 
because it displays its diffraction colors, "play of color," not like precious opal by il- 
luminating and observing its face, but by passing light through the stone. A weak 
expression of this characteristic of contra luz opal can be seen in a small percentage 
of the Mexican opal known as hyalite opal when a small chunk, especially if wet, is 
held toward a light. Hyalite opal is defined in the Funk and Wagnalls Dictionary as 
"a pellucid glassy variety of opal of no commercial value." 

This author has collected a considerable number of pieces of such opal because 
it is important in unravelling the puzzle of the natural formation of opal (1, 2, 3). In 
cutting small slices off the rough faces of such opal, in order to see the structural 
features within, vivid interference colors were encountered in perhaps half of the 
hyalite opal nodules. These vivid colors were visible only in thin slices. Cursory ex- 
amination of this opal suggested that the colors may have an origin similar to that 
of play of color in precious opal. 

The structure of precious opal which gives rise to characteristic brilliant and 
pure interference colors was elucidated by a group of collaborating Australian 
scientists: Darragh, Gaskins and Sanders (2), who employed scanning electron 
microscopy to show that precious opal contains a regular lattice of closely packed 
silica spheres of uniform size and spacing, appropriate to cause color separation of 
visible light by optical interference. 

The distance between layers in the lattice, d-values, were found to be approx- 
imately one half the wave length of visible light in a medium of refractive index of 
opal, 1.45. The dimensions found also correlated with the dimensions required to 
produce the predominant wave length of the play-of-color of different stones. The 
change of color with changing angle of illumination and observation correlated well 
with the expectations for such a diffracting system. They also studied Mexican 
precious opal which occurs in cavities of igneous rocks and an Australian opal of 
similar form. Using scanning electron microscopy, they studied such opal only 
enough to establish that these opals also contain a regular lattice structure of 
similar dimensions. 

To my knowledge, no comparable study has been reported on the structure or 
optical properties of contra luz opal, perhaps because of its relative non importance 
as a gem stone. Interest in this stone also has been limited by the failure to 
recognize the importance of thin slicing. The facility of an electron microscope has 
not been available to me. It was, therefore, decided to determine if the observed 
play of color and optical properties could be explained on the basis of a regular lat- 
tice structure by analogy to the understanding of the structure of precious opal. 

Observations 

For comparison purposes, several Australian precious opals were sliced flat 
and the face ground flat by lapping in a direction and at a level to coincide with a 
layer of play of color in its face. After grinding with a 600 mesh diamond lap, a 

368 



Physics and Astronomy 



369 



microscope cover slip was cemented over the flat face with epoxy 330 cement. Such 
flat faces simulate a well polished flat face. 

These faces were illuminated with near parallel white light and observed and 
photographed at various angles. As the angle between illumination and viewing in- 
creases, the interference colors move to shorter wavelengths. For instance, an area 
which is green when illuminated and viewed at 10° from normal to the surface is 
blue at 30° and colorless at 70°. Figure 1 shows the geometry of this arrangement. 

Representative thin slices of contra luz opal, generally less than 1 mm thick, 
were cut from hyalite opal nodules. Unlike the situation with Australian precious 
opal, the hyalite opal generally was sliced in a vertical orientation, that orientation 
being evident by the existence of a natural flat top on the nodule. These slices also 
were ground flat on a diamond lap and had a microscope slide and a cover slip 
cemented with epoxy 330 onto their faces. This results in very little scattering at 
the faces. 

Optical arrangements were made to view and photograph the slices at varying 
angles when illuminated with a parallel beam of high temperature incandescent 
light. The observed colors against-the-light contrast with a total absence of play of 
color when these slices are observed under face illumination even under ideal con- 
ditions, for instance, when they are backed with a black plate. The observation and 
photography of these contra luz slices were made by the geometry shown in Figure 
2. It is to be noted that the angle between the incident beam and the diffracted ray 
-within the stone - is much greater than the maximum angle achievable between in- 
cident light and diffracted rays in the system shown in Figure 1. 

The play of color seen in such slices is generally less well separated into zones 
of a single color than is seen generally in Australian precious opal. The colors 
generally stream or grade into one another. It may well be that this distinction 
would be less prominent if the contra luz opal had been sliced generally along the 
horizontal direction as is generally done with Australian precious opal. 

The play of color in a slice of contra luz opal shifts to shorter wavelength, en- 
tirely as expected, as the angle between the incident beam and the diffracted rays 
increases in these experiments. This is documented in photographs which can not 
be reproduced in this report. When the colors seen at specific angles between the 
incident and observed directions were employed to calculate d-values, the values 
were, of course, much greater than those calculated from observations on precious 
opal. Colors all the way from red to blue are seen in contra luz opal. 





FIGURE 1— a— Path of a ray entering or leaving opal, n = 1.45 
b— Scheme 1 face illumination (for precious opal) 



370 



Indiana Academy of Science 





Figure 2 — Scheme II against-the-light viewing (for contra luz opal) 





Figure 3 — Scheme HI side light illumination (for medi opal). 



Some of the slices of hyalite opal prepared as above were found to produce lit- 
tle play of color when illuminated as in Figure 2. They were seen to: 1) show more 
color when illuminated at minimum angles in scheme 2; and 2) show intense play of 
color when the light was allowed to enter the edge of the stone and the face of the 
stone was observed. Analysis of this situation led to the understanding of the 
scheme of observation shown in Figure 3 and discussed in the later section. 

Optical Considerations 

When light enters a transparent solid from air at any angle other than normal 
to the surface, it is bent by an angle which depends on the refractive index of the 
solid and the incident angle. The refractive index of air is approximately 1.00 and 
that of opal is 1.45. The relation of the internal angle to the external angle is given 
by the formula sin a = n s where a is the angle between the incident direction in air 

sin b 
and the normal to the surface, b is the corresponding angle between the incident 
ray inside the stone and an extension of the normal to the surface into the solid, and 
n g is the refractive index of the solid, here 1.45. When a ray leaves the solid and 
enters air, it is bent identically at the surface, a and b having the same meaning as 
above. Figure 1 portrays the path of an incident ray i and that of a diffracted ray d 
which originates at a point L in the solid. In the first scheme of observation of in- 
terference colors used in viewing precious opal, this geometry is employed. 

In the present treatment, the angles for entering and for emerging rays are 
kept equal to one another for simplicity of treatment. This restriction on viewing 
the stone does not limit the arguments which follow. 

In the simplest case of illuminating and viewing the stone, both at normal 



Physics and Astronomy 371 

angle to the face where a = (and b = 0) the relation of d, the distance between 
plates or layers of a diffracting system, to the wavelength of light giving construc- 
tive interference (X air and X opa j) is simply: 

,» , opal *air 

1) d = 



2 1.45 x 2 

Since diffraction occurs inside the stone, the angles between the incident and 
diffracted rays inside the stone are used for calculations. One half the angle be- 
tween incident direction and the direction of the diffracted ray is designated as (3 in 
all schemes of observation. 

In the more general case where the angle (3 is anything between 0° and 43.5°, 
the theoretical maximum value corresponding to a = 90°, the relationship of d, the 
angle 0, and the wavelength of the diffracted ray in air or opal is given by the ex- 
pressions which hold also for Schemes 2 and 3. 

\ 

1.45 



Val = 2d bVfl- tan/3sin/3 ) 



X air 



2x 1.45 (^jg-tan/3 sin j3j 

Table 1 contains the results of calculations of the d-values for typical in- 
terference colors when incident and diffracted rays are assumed to take several 
values of a and the consequent values of (S. Note that the values at a = 0° and a = 
90° are theoretical limits. Feasible limits of observation are more nearly a = 20° 
and a = 70°. Calculated d-values for these angles are also given in Table 1. This 
scheme of viewing is applicable to precious opal and is referred to as scheme 1. 

The geometry of observations of contra luz opal - by looking through a thin 
slice of stone - is illustrated in Figure 2. In this case the angle between the incident 
and diffracted rays within the stone is greater than in scheme 1, having values be- 
tween 2 x 46.5° and 2 x 90°. (3 the half angle, is between 46.5° and 90°. The feasible 
limits on viewing angles a are approximately 70° and 20°, corresponding to angles 
(3 = 49.6° and 76.4° respectively. Thus neither the feasible nor the theoretical 
values of /3 for schemes 2 and 1 overlap. 

Table 1 contains the results of calculations of d-values for contra luz opal 
showing interference colors at feasible angles of observation. The theoretical 
minimum angle (3 is 46.5° and the theoretical maximum of (3 is 90°. But at (3 = 90° 
the value of d is infinite for all wavelengths and is not significant. Since the (3 
ranges for schemes 1 and 2 do not overlap no lattice giving interference color by 
scheme 1 viewing can give the same color by scheme 2 viewing, and the reverse is 
true. 

Of course a given stone might have lattices of widely varying d-values at dif- 
ferent places. Different colors exhibited at different locations in a given stone at 
one angle of observation are an indication of this fact. In gemology a precious opal 
exhibiting different areas of color from red to blue at one angle is exceptional and is 
most highly prized. 

If an opal had a lattice with a spacing d between the feasible ranges for 
precious and contra luz opals, it would not show interference colors by these 
schemes of viewing. Fortunately, a third scheme of viewing was discovered as 



372 



Indiana Academy of Science 



a 






2 '% 



g s 



* > 



£ J3 



^J£ 



5 " gig 
s: <a Halm 

S3 



ID 4) 
5 O 



Physics and Astronomy 373 

referred to earlier. We shall call it scheme 3 and represent it as in Figure 3. This 
scheme permits observation at angles intermediate between and overlapping 
those of scheme 1 and scheme 3. The observation of vivid interference colors by 
this scheme, which may be referred to as "side light illumination," evidences con- 
tinuity of d-values in ordered lattices through the range between those of precious 
opal and contra luz opal. 

In scheme 3, the slice is illuminated through its edge and viewed through its 
face. In this case /3 has feasible values from 29° to 60.5°, corresponding to viewing 
angles between about 50° on each side of the normal to the face. Table 2 contains 
the results of calculating d-values for various colors viewed at feasible viewing 
angles relative to the incident direction by scheme 3. The author suggests the 
name medi opal for Mr. Opal of intermediate d-volumes. 

Conclusions 

These observations on three classes of opal support the conclusion that the 
sizes of silica spheres which form regular lattices in opal capable of producing in- 
terference colors vary widely outside those sizes observed by Darragh and 
coworkers for precious opal, and extend to at least double the diameter and inter- 
layer spacing for precious opal. An extension of the electron microscope studies of 
Darragh et al. to hyalite opal would be pertinent. 

While no statistical study was made of the distribution of d-values in a signifi- 
cant number of hyalite opal nodules, the spread of values in ten nodules ranged 
from approximately 300 nm to 800 nm. It may be supposed that opals do exist hav- 
ing d-values higher than those which show interference at feasible high angles of 
viewing by scheme 2. The highest d-values which one would be able to detect by 
this kind of viewing would give a blue interference near a = 0° in scheme 2. 

To test this idea, an optical system is being put together to extend the feasible 
limit of observation to (3 values close to 90° in scheme 2. The upper limit of d-values 
which could be determined in this way depends only on how close to (3 = 90° one 
can make observations. 

Perhaps the most significant question raised by these conclusions is: What 
process of natural formation leads to such wide ranging d-values, notwithstanding 
the regularity of the lattice required for the production of interference colors? This 
question was discussed by the author in reference 3. 

Table 2: Calculated Lattice d-Values for Interference Colors at Feasible Angles 
for Medi-oPal (side lighted) 





(nm) 
IN AIR 


Wavelength 


MEDI-OPAL AT ANGLE 




COLORS 


a* 0** 
-50 29° 


a 
45° 


a 
+ 50 60.5° 


Blue 


470 


185 


229 


333 


Green 


520 


204 


253 


368 


Yellow 


580 


228 


282 


410 


Orange 


600 


236 


292 


425 


Red 


650 


256 


317 


460 



* a is the external angle between the normal to the surface and the viewing direction. This angle is measured 
in a plane including the direction of the incident light. It is considered negative when it makes an acute angle 
and positive when it makes an obtuse angle with the direction of the incident light. See FIGURE 3. 

•• is here: = (45°--|) and = (45° + y) 



374 Indiana Academy of Science 

Literature Cited 

1. Darragh. P. J. and A. J. Gaskin. 1966. The Nature and Origin of Opal. 
Australian Gemmologist #66 p. 5-9. < 

2. Darragh, P. J., A. J. Gaskin and J. V. Sanders. 1976. Opals. Scientific 
American 234 #4 p. 84-85. 

3. Davis, W. W., Indiana Academy of Science, Proceedings. 1978. p. 237-^41. 



Microcomputer Assisted Multiple Coulomb 

Scattering Measurements in Nuclear 

Emulsion Pellicles 



Gerald P. Thomas and Mark Coggins 
Department of Physics and Astronomy 

Ball State University 

Muncie, Indiana 47306 

Introduction 

Attempts have been made in the past to automate various aspects of 
microscope measurement of elementary particle tracks in nuclear emulsion targets 
exposed to high energy primary particle beams from particle accelerators. These 
attempts, based largely on obtaining magnified, external 2-d projections, have 
proved to be uncompetitive since they do not incorporate the built-in 3-d "stereo" 
view advantage inherent to the microscope method of measuring collision star 
tracks. However, there is great labor in performing measurements partaking of 
this "human" factor. Recently, however, the use of on-line, inexpensive microcom- 
puters can simplify and improve the taking of data during various measurement 
algorithms as well as provide immediate feedback of measurement results toward 
on-line re-measurement decision making. 

In this work, we will discuss the adaptation of a KIM-1 (6502) based microcom- 
puter to the automation of the technique of "multiple scattering" that determines 
particle track momentum from measurements of the "waviness" of the track of 
grains. 

Method 

A schematic of the automation architecture hardware used is seen in (Figure 
1). In a typical "multiple scattering" measurement involving the y-coordinate, the 
Koristka measuring microscope's micrometer eyepiece scale moves up and down 
relative to a stationary notch as the external knob shaft is rotated clockwise or 
counterclockwise respectively, thus indicating a change in the crosshair's 
y-coordinate. 

A Dynamics Corporation Model 77 optical encoder, mounted concentric to the 
knob shaft, transduces a y-coordinate "first difference", amounting to two shaft 
rotations, into a total of 2,880 successive square pulses (5 volts each). At 550X 
magnification, this amounts to 4.66 microns in the emulsion. Actually, two parallel 
pulse trains A & B are sent out, A leading B by 90° for a clockwise rotation and B 
leading A for a counterclockwise rotation. 

On the processing board is a logic circuit shown in (Figure 2) that gates 
through the first train of pulses (A or B) that arrive there and suppresses the trail- 
ing train. 

The board also contains a NOR gate (see Figure 3) whose inputs are tied to the 
A & B input lines which enter the computer port, so that its output goes low every 
time a pulse of the surviving pulse train (A' or B') arrives at the port bits. Since the 
gate's output is tied to the maskable interupt (IRQ), an A or B pulse signals the 
computer to proceed to either the add or subtract service machine subroutines (see 
KDPASM below). The output of the NOR gate is also tied to still a third port bit, so 

375 



376 



Indiana Academy of Science 




"J Processing Board 



Microcomputer (KIM- 1) 



Terminal 



Switch 



mamscale 



Modem j 



Microcomputer (DEC-I) 




Figure 1. Eyepiece Scale Automation Architecture 

the software can determine when each pulse is finished in order to set up for the 
next pulse arrival. 

A machine language program, previously read in, is used by the KIM-1 to pro- 
cess the various pulse totals for microscope knob twists. This three page (% K, 
RAM) measurement program (KDPASM) is discussed below. A pedal is tied 
through a NAND gate debouncing circuit on the board to the non-maskable (NMD 
interrupt and is used to jump to various interrupt subroutines of the program dur- 
ing measurement. 

By switching the KIM-1 into the role of a terminal for the macrocomputer 
(DEC-10), the data that was collected and collated by the KIM-1 program can then 
be dumped through a modem and phone line to the large computer (DEC-10) for 
computation of particle momentum using the FORTRAN momentum program 
(SCAT). The function of the terminal (DEC WRITER) is to open and close the hex 
data input file, and request execution of the SCAT momentum program and finally 
display the momentum results leading to possible re-measurement. 




fCH> 



A' (if A leads B) 



'^CM> 



B'(if B leads A) 



Figure 2. Angular Direction Logic Circuit 



Physics and Astronomy 



377 



PORT 



A' 



B' 



^> 



to IRQ 

Figure 3. Data Aquisition at Port 



The switch (see Figure 1 and Figure 4) permits 1) half-duplex between the 
KIM-1 and the DEC-10 for dumping data to the DEC-1 data file and 2) full-duplex 



r^Wn 



© to © 




Moke- Before- Break 
Switch 



Figure 4. Associated Switch Circuitry 



378 



Indiana Academy of Science 



between the terminal and the DEC-1 for setting up data files and calling fortran 
programs in the user disk file-in one direction, and receiving calculation results on 
the terminal-in the other direction. 

An associated 7422 opto-isolater chip and a make-before-break feature insures 
that the DEC-10 does not break off its line of communication with either the KIM-1 
or the terminal during switch over between them as inputs devices to the DEC-10, 
and circumvents electrical ground ambiguity. 

The software machine program (KDPASM) was written to receive groups of 
pulses such as are generated during a typical "multiple scattering" (1) measure- 
ment as depicted in (Figure 5). Now, once demographic parameter data is keypad 
entered, several sets of pulses are generated, one for each cell length of x-motion in 
the general direction of the track. 

The overall program description and flow diagram is seen in (Figure 6). A 
pedal interrupt (NMD signals the end of each segment (of one cell length) and 2 
pedal interrupts signal either the end of a track or the end of the "star" of tracks as 
indicated by pressing the D (dump) key as shown. 

Note that within the NEXTSEG subroutine, the segment's net up ( + y) or 
down ( - y) (clockwise or counterclockwise knob rotation respectively) number of 
pulses is always refered to a value of 8000jj (reset for each segment of track). For 
example, an outputed value of 7F8Fjj means 8000tt-7F8Ftt = H^iq net 
counterclockwise pulses have been recorded for a segment and 82FOj_j means 
82FOh - 8000 H = 02FOh = 752 10 net clockwise pulses. 

A sample DEC-10 data file printout, seen exactly as stored and outputed from 
the KIM-1 in hex form, is shown in (Table 1) and in decimal form suitable for data 
input to SCAT in (Table 2). The pulses per segment are converted to main scale 
divisions (Ay) per segment by dividing by 2880 (2880 pulses = 1 eyepiece division) 
and then microns by multiplying by 4.66 microns per division. This is done in the 
SCAT fortran program. 

The multiple scattering of a charged elementary particle due to collision with 
emulsion nuclei coulomb fields gives rise to (1) an observed average second dif- 
ference <D> of y-readings that is related to the momentum p as follows (1): 



Crosshairs 




Notch Scale 



Vertex 



Field of 
View 




Track of 
Grains 



1 (eyepiece 
<4s scale 
I direction) 



Figure 5. Multiple Scattering Measurement Procedure 



Physics and Astronomy 



379 



MAIN (Initialize and Read in Event Parameters) 




Read in parameters 
of next track of event) 



(idd or subtract 
sucessive encoder 
pulses over one 
segment) 



( record total pulse 

count for each 
segment) 



(either go to next 
track in event or 
go to event DUMP 



KEY (D) 



DUMP 



(dump parameters 

and encoder data 
to DEC file 

8 
then go to start 

of next event 



Figure 6. KDPASM Machine Program Flow Diagram 



Table 1. Hex Data File 



Name 
AA 



Plate 
101279 



Plate 
22 



Type 
0003 



Event 
0071 



Microns/eyepiece scale division 
0466 



01 — One track in event 

01 — Track number one 

0250 -Cell length 

06 — Twice number of segments 



3 Segments 



7F8F-113 Pulses Counterclockwise (below 8000) 
814C-332 Pulses clockwise (above 80001 

82FO-752 Pulses clockwise (above 80001 



380 



Indiana Academy of Science 
Table 2. Decimal Data File 



Name Event 

AA 71 



250 
cell length 



Type 
00 + 03P 



500 
2X cell length 



Date Name 

101279 AA 



scale 



04 
readings 



track number 



10,000 



09887 



10219 



10971 



113 pulses + 332 pulses 752 pulses 



P0C 



KC 



573<D> 



— (p in Mev/c) 



Where 

K = approximate constant from theory ( = 47) 

C = cell length in microns 

<D> = average second difference 

13 = v/c 

v = particle speed 

c = light speed 

This is computed by the FORTRAN program (SCAT) and returned to the terminal. 

The overall sequence of software relevant to the multiple scattering 
algorithm for finding particle momentum then consists of using the machine file 
KDPASM to receive, treat and store encoder pulse data and then dumping the hex 
data to the DEC-10 macrocomputer, once a data file has been opened from the ter- 
minal. After receipt of data in this file, the file is closed and a fortran program 
(HEXCVT) located in the DEC-10 converts the hex format to a decimal format. This 
decimal data file is then used by the SCAT fortran program to compute and return 
the sought after momentum to the terminal. 

Results and Conclusions 

The KIM-1 microcomputer based automation proved to be adequate to pro- 
vide relatively immediate feedback of particle momenta following measurement of 
eyepiece scale y-coordinate displacements. In particular, of crucial importance was 
the relatively short KIM-1 (6502) word-time coupled with short enough add & sub- 
tract software routines. This allowed the microprocessor to not miss any pulses ar- 
riving at the port during real time "counter" activity. Use of the DEC-10 to make 
computations is somewhat inconvenient in the sense of manual switching required, 
DEC-10 down times, etc., but computation time is short with the DEC-10, and only 
modest KIM-1 RAM memory is required for both machine programming of data 
manipulation (% K) and for data storage (few K). With the current influx of not too 
expensive 16 bit, higher speed microcomputers, it would be possible to time-share 
real-time service of encoders attached to all three microscope dimensions. This 
would be a giant step toward cheap automation of coordinate measurement both in 
scanning and plotting of collision events and in multidimensional measurement pro- 
cesses in the emulsion detection approach to High Energy Physics. 



Physics and Astronomy 381 

Literature Cited 

1. Powell, C. F., P. H. Fowler and D. H. Perkins. 1959. The Study of Elemen 
tary Particles by the Photographic Method. Pergamon Press. London. 
114-129. 669 pp. 



PLANT TAXONOMY 

Chairman: John A. Bacone 

Director, Division of Nature Preserves, Indiana Department of Natural Resources, 

Indianapolis, Indiana 46204 

Chairman-Elect: Richard J. Jensen 
Department of Biology, St. Mary's College, Notre Dame, Indiana 46556 

ABSTRACTS 

The Flora of the Southeastern United States: A Review. Buddy Cantrell, 
Department of Biology, The University of Notre Dame, Notre Dame, Indiana 

46556. Beginning with the earliest expeditions to the New World, the flora of 

the North American colonies was of great interest to botanists and hor- 
ticulturalists, alike, in Europe. The travels, collections, and correspondence of 
early biologists including John Banister and John and William Bartram introduced 
much of the European community to the southeastern flora. The notes and 
specimens of John Clayton served as the basis of Gronovius' Flora Virginica and 
southern plants, particularly from Virginia and the Carolinas, also were reported in 
works by such Old World masters as Linnaeus and Andre Michaux. A prominent 
interest in the flora of North and South Carolina was first evidenced by Thomas 
Walter's Flora Caroliniana and, later, by Stephen Elliot's Sketches of the Botany 
of South Carolina and Georgia. The first regional flora was Flora of the Southern 
United States by Alvan Wentworth Chapman and the first state floras were 
Augustin Gattinger's Flora of Tennessee and Charles T. Mohr's Plant Life of 
Alabama. John K. Small published works concerning the flora of Florida and pro- 
duced three regional treatises. His Manual of the Southeastern Flora is the most 
recent, comprehensive flora though several excellent state and specialized 
treatments have been completed. Continuing biosystematic and floristic work by 
southeastern botanists has, in part, satisfied the floristic needs of their respective 
interests and has produced smaller works of great interest to the taxonomic com- 
munity. These endeavors provide the framework for the Vascular Flora of the 
Southeastern United States of which the first volume, Asteraceae by Arthur Cron- 
quist, was released in 1979. 

An Updated, Computer-based Checklist Of The Vascular Plants of Indiana: The 
Unending Synthesis. Theodore J. Crovello, Department of Biology, The Univer- 
sity of Notre Dame, Notre Dame, Indiana 46556, and Clifton Keller, Andrews 

University, Berrien Springs, Michigan. The flora and fauna of any geographic 

area may seem fixed and unchanging, but it is actually in dynamic and constant 
change. For an area the size of Indiana, this change is caused by: habitat modifica- 
tions (both natural as well as human caused); by inherent characteristics of 
organisms; and by the amount of taxonomic and biogeographic knowledge available 
at a given time. This last factor affects what taxa are considered to be present in 
Indiana, but it also mandates changes in taxonomic nomenclature. This "unending 
synthesis" also mandates changes in taxonomic nomenclature. This "unending syn- 
thesis" of knowledge and resultant nomenclatural changes causes problems for 
people in both academic and applied positions. For example, use of Deam's 1940 
Flora of Indiana does not reflect either nomenclatural changes of the taxa he 
described or the addition of the new taxa discovered growing in the state since 
1940. We have developed a computer-based checklist of the vascular plants of In- 

382 



Plant Taxonomy 383 

diana. Nomenclatural changes are based on the recently published checklist of 
North America, North of Mexico by J. T. and R. Kartesz. Sources of information on 
taxa new to the state included: those published in the Proceedings of The Indiana 
Academy Of Science since 1940; those in Swink's 1974 edition of the Flora of the 
Chicago Region; and the Indiana Department of Natural Resources (particularly 
the Division of Nature Preserves), Our computer-based checklist allows us to relate 
any nomenclatural changes since Deam with the names that he used. This linking of 
the most recent nomenclature with that use in the standard and classic floristic 
work of Indiana (Deam's 1940 Flora) was a high priority. The checklist not only will 
be available in regular published form, but also as a customized computer printout 
that will include the latest information on new state records and nomenclatural 
changes. Readers are requested to send fully documented herbarium specimens of 
possibly new state records to Crovello at the University of Notre Dame to assure 
proper credit and inclusion in the checklist. 

Phylogenetic Reconstruction in Quercus. Richard J. Jensen, Saint Mary's College 

Notre Dame, Indiana 46556. Fifteen taxa of red oaks native to the eastern 

United States were examined with the objective of estimating phylogenetic rela- 
tionships within the group. A set of 28 characters was used as the basis for 
UPGMA cluster analysis, Wagner Tree construction, and character compatibility 
analysis. The phenetic and compatibility tree diagrams were used as input to the 
Wagner algorithm and lengths and deviation ratios were determined for each. 
Standard Wagner analysis produced two trees: a tree of length 107 was produced 
with the OTUs arranged alphabetically and a tree of length 106 was produced after 
the OTU sequence was randomized. The lengths of the phenetic trees varied from 
110 to 114, a tree approximating Trelease's 1924 view of species relationships had a 
length of 120, and the compatibility tree had a length of 111. Deviation ratios were 
lowest for trees based on compatibility (0.55), correlations from standardized 
characters (0.56), distances from standardized characters (0.58), the Wagner Tree 
of length 107 (0.59), and the Wagner Tree of length 106 (0.61). Perhaps the most in- 
teresting aspect of the study is revealed by comparing relationships depicted by 
these trees with those illustrated by two independent data sets. In both cases, the 
tree exhibiting the greatest agreement with these new data sets is that produced 
by character compatibility. Even so, there are reasons to question all of the derived 
trees. The basic problems seem to center on (1) the possibility of the group being 
polyphyletic, (2) the possibility of misrepresentation of character trends, and (3) the 
possibility of reticulate evolution. 

Vascular Plants of Sand Hill Nature Preserve, Pulaski County, Indiana. Victor L. 
Riemenschneider, Department of Biological Sciences, Indiana University at South 

Bend, South Bend, Indiana 46615. Sand Hill Nature Preserve, at 24.3 ha area, is 

located in the northern one third of Tippecanoe River State Park, 11.3 km north of 
Winamac, Indiana. The preserve and surrounding area are within the Outwash and 
Dune Facies of the Atherton Formation. The soils are medium to strongly acid, 
very poorly to excessively drained and sandy textured. The pre-settlement vegeta- 
tion of this area ranged from oak forests dominated by white and black oak to wet 
marsh prairie. Today, the preserve communities are black oak forest, pin oak-black 
gum-red maple swamp forest, scotch and white pine plantations and degenerate 
shrub prairie. The 220 species of vascular plants identified in the preserve area 
reflect the vegetational history of the preserve including early attempts to farm 
portions of the area. None of the species are on Indiana's special plants list. An 
alphabetical list of species is available from the author. 



384 Indiana Academy of Science 

The Epidermal Anatomy of Deeringothamnus Small and Its Taxonomic Implica- 
tions. John L. Roth, Jr. and David L. Dilcher, Department of Biology, Indiana 
University, Bloomington, Indiana 47405. The generic status of Deeringotham- 
nus Small has been argued extensively in systematic literature for the past fifty 
years. Some favor retaining Deeringothamnus as a separate genus while others 
would combine it with Asimina Adans. However, due to a lack of compelling 
evidence the systematic treatment of Deeringothamnus has been largely a matter 
of personal taxonomic judgment. This report presents for the first time a 
characteristic of the two Deeringothamnus species which is unique among the 
more than 2000 species of the Annonaceae. The leaves of Deeringothamnus are am- 
phistomatic: stomatal complexes are found on both the abaxial and adaxial sur- 
faces. Together with evidence pointed out by previous authors, especially the dif- 
ferences in pollen morphology, this character argues for the recognition of Deer- 
ingothamnus as a separate genus of the Annonaceae that is closely related to but 
distinct from Asimina. 

Najas marina L. in Indiana. Helene Starcs, Indianapolis. Najas marina L., the 

spiny naiad, was found by the author growing in the highly calcareous Beaverdam 
Lake, Steuben County: 8-26-1974 and 8-31-1980. This is a new plant distribution 
record for the state of Indiana. 






The Status of Indiana's Rarest Plants: 
A Revision of the List of Endangered and Threatened Vascular Plants 

John A. Bacone and Cloyce L. Hedge, 

Indiana Department of Natural Resources, 

Indianapolis, Indiana 46204 

and 

Theodore J. Crovello, 

University of Notre Dame, 

Notre Dame, Indiana 46556 

Introduction 

The purpose of the study was to update the knowledge concering Indiana's 
rarest plants, especially those which were being considered by the United States 
Fish and Wildlife Service for federal endangered and threatened status. It was 
hoped that the data to be collected would aid in ascertaining the true status of all of 
Indiana's rarest plants. All data collected were to be used to keep the data bases of 
the Biology Survey Committee of the Academy and the Indiana National Heritage 
Program up-to-date. 

Methods 

The staffs of the Biology Survey Committee, the Division of Nature 
Preserves, and the Natural Heritage Program all were involved in the preparation 
and functioning of the study. The state was divided into five regions, and several 
regions were split into two subregions, based upon size and number of sites to be 
checked. Field botanists were hired to work in six regions or subregions. These 
were supplemented by the efforts of the Hayes Arboretum, who worked in 
southeast Indiana, and by Dr. Richard Maxwell, Indiana University Southeast, who 
worked in Clark and Floyd Counties. Department staff worked in central Indiana 
counties. 

Information was provided to the botanists monthly, on a computer printout 
provided by the Heritage Program. This included all information gleaned from her- 
barium labels, and as exact a location as possible. The information was provided for 
all plants included on the "Preliminary List of Endangered and Threatened 
Vascular Plants of Indiana", by Bacone and Hedge (1). Priorities were given to each 
plant site, with those plants of federal concern, extirpated or endangered status 
being the top priority for field checking. A set of topographic maps was also pro- 
vided. 

The botanists were asked to check as many of the top priority sites as possible 
each month, and were expected to work six days a month from May through 
September. The botanists provided information as to condition of habitat if the 
plant was not found. If the plant were found, information collected included exact 
location, habitat, threats, population size and viability, and associated species. 
Whenever possible, especially in cases where identification was difficult, a slide or 
voucher specimen was collected, unless such collection would seriously jeapordize 
the population. 

Results 

During the field season, attempts were made to reverify 887 sites of rare 

385 



386 Indiana Academy of Science 

plants. Of these, 280 were found, or a 32% "success" rate. Two hundred and 
twenty-three plants were searched for with "endangered" status, and sixty-one 
were re verified (27%), twenty-eight reverified of fifty-eight with federal status 
(48%), and eighty-four of 275 with threatened status (31%). The other plants sear- 
ched for had a "rare" status, or another status of "special concern". In addition to 
the reverifications, a number of new locations were discovered. Overall, 265 new 
sites were found, including thirty-five (federal status), 47 (endangered status), 76 
(threatened status), and 74 (other categories). Approximately 140 sites, 16% of 
those looked for, were considered destroyed. The remainder were not located but 
the habitat appeared to be extant, or the locations were too vague to be checked. 

As a result of this field work, status changes have been proposed for several 
species. These are listed with the original status (from Bacone and Hedge (D) 
shown in parentheses, followed by the revised status. Nomenclature follows 
Gleason and Cronquist (2). 

Anemone caroliniana Walt. (Endangered) Extirpated 
Antennaria solitaria Rydb. (Threatened) Special Concern — Rare 
Aristolochia tomentosa Sims. (Threatened) Special Concern — Rare 
Betula populifolia Marsh. (Endangered) Extirpated 
Habenaria leucophaea (Nutt.) Bray. (Endangered) Extirpated 
Hypericum dolabriforme Vent. (Extirpated) Endangered 
Hypericum frondosum Michx. (Endangered) Extirpated 
Ilex decidua Walt. (Threatened) Special Concern — Rare 
Illiamna remota Greene. (Special Concern — Federal) Endangered 
Iresine rhizomatosa Standi. (Endangered) Threatened 

Lesquerella globosa (Desv.) Wats. (Endangered) Special Concern — Taxonomy 
Melica mutica Walt. (Threatened) Special Concern — Rare 
Perideridia americana (Nutt.) Reichenb. (Threatened) Endangered 
Phyllitis scolopendrium (L.) Newm. var. americana Fern. (Special Concern — Federal) 
Dropped 

Polytaenia nuttallii DC. (Threatened) Endangered 
Rudbeckia fulgida Ait. (Threatened) Special Concern — Taxonomy 
Rudbeckia fulgida Ait. var. umbrosa (Boynton and Beadle) Cronq. (Endangered) 
Special Concern Taxonomy 

Rudbeckia palustris Eggert. (Endangered) Special Concern — Taxonomy 
Saxifraga forbesii Vasey. (Special Concern — Verification) Endangered 
Scutellaria parvula Michx. var. australis Fassett. (Threatened) Special Concern — 
Taxonomy 

Sparganium androcladum (Engelm.) Morong. (Endangered) Special Concern — Tax- 
onomy 

Stachys clingmanii Small. (Threatened) Endangered 
Wisteria macrostachya Nutt. (Endangered) Extirpated 
Zygadenus glaucus Nutt. (Endangered) Threatened 

Discussion 

The Department of Natural Resources and the Academy of Science continue 
to be interested in ascertaining the true status of Indiana's rarest plants and 
animals, and hope to continue with similar field work in the coming years. This 
study has provided much needed information, which can be used to help protect the 
rarest elements of our flora, through a combination of tools such as the planning 
and review process, acquisition, registration as a natural area, and research. It is 
hoped that all persons knowledgeable about any of these plants will continue to 



Plant Taxonomy 387 

communicate concerning threats, new locations, status information, etc., to both 
the Academy and the Department, so that the information base can be as up-to- 
date as possible. Continuing field work such as this effort are desperately needed in 
the years ahead. 

Acknowledgments 

The study was made possible in part through a grant from the United States 
Fish and Wildlife Service to the Biology Survey Committee of the Indiana 
Academy of Science. Staffs of the Division of Nature Preserves and the Indiana 
Natural Heritage Program of the Department of Natural Resources provided 
valuable assistance. We are especially grateful to the field botanists who worked 
on this project: Kenneth Dritz, Cloyce Hedge, Larry Hauser, Dr. John Ebinger, 
Maryanne Newsom-Brighton, Lois Mittino Gray, and Dr. Richard Maxwell. Special 
thanks to Helene Stares for aid in species identification and to Jim Aldrich for pro- 
viding computerized information. Thanks are also extended to the Hayes Ar- 
boretum for funding the field work completed by Don and Elaine Hendricks. 

Literature Cited 

1. Bacone, J.A. and C. L. Hedge. 1979. A preliminary list of endangered and 
threatened vascular plants in Indiana. Proc. Ind. Acad. Sci. 89:359-371. 

2. Gleason, H. A. and A. Cronquist. 1963. Manual of vascular plants of nor- 
theastern United States and adjacent Canada. D. Van Nostrand Co., 
Princeton, New Jersey 810 p. 



Plant Distribution Records for Rush, Shelby and Decatur Counties 

Mary Patricia Coons 

Department of Biology, Indiana University 

Bloomington, Indiana 47406 

In preparation of his Flora of Indiana, Deam (1) obviously was unable to collect 
with equal thoroughness in all counties of the state. Rush, Shelby and Decatur 
Counties are three adjacent, largely agricultural counties of central Indiana where 
collection was less intense and relatively little subsequent work has been done. The 
low totals for species reported for these counties shown in the map presented by 
Keller (3) reflect a lack of collecting activity rather than depauperate floras. 

During 1979, the author had an opportunity to collect plants in these counties, 
and the new county records are reported here. Plants are listed according to county, 
with the genera arranged in the order of their appearance in Deam (1) and species 
alphabetized within their respective genera. Nomenclature is according to that 
used in Gray's Manual of Botany, 8th ed. (2). Vouchers of all species listed are 
deposited in the herbarium of Indiana University. A. H. and Alvene Coons assisted 
in the collection of some specimens and the preparation of the lists. 

Most of the species included are fairly common and widely distributed in the 
state; however, a few deserve special comment. Ampelamus albidus (Asclepiada- 
ceae) has become widespread and common in the state since Deam's time. The 
weedy umbellifer Torilis japonica, formerly reported from a rather limited area in 
central Indiana, appears to have extended its range significantly and may, in fact, 
have become fairly common while remaining unnoticed because of its superficial 
resemblance to Daucus Carota and its occurrence in similar habitats, mostly road- 
sides and edges of fields. Collections of Cirsium arvense (Compositae) help to docu- 
ment the recent outbreak of that weedy species in central Indiana. Tragopogon 
porrifolius (Compositae) was found in Rush County in a mixed population with T. 
pratensis and their hybrid. 

Taxonomic Entities 

Decatur County: Hemerocallis fulva, Tklaspi perfoliatum, Hesperis 
matronalis, Medicago lupulina, Melilotus alba, Melilotus officinalis, Coronilla 
varia, Apocynum cannabinum var. cannabinum, Asclepias syriaca, Ampelamus 
albidus, Glechoma hederacea. Prunella vulgaris, Verbascum thapsus, Achillea 
millefolium, Chrysanthemum Leucanthemum var. pinnatifidum, Cirsium arvense. 

Rush County: Equisetum arvense, Equisetum hyemale var. affine, Hordeum 
jubatum, Hemerocallis fulva, Allium vineale, Trillium fie xipes, Trillium sessile, 
Carya ovata, Ulmus americana, Polygonum Convolvulus, Stellaria pubera, 
Isopyrum biternatum, Ranunculus repens var. repens, Ranunculus septen- 
trionalis, Dicentra canadensis, Brassica nigra, Capsella Bursa-pastoris, Draba 
verna, Hesperis matronalis, Sedum tematum, Potentilla recta, Rosa palustris, 
Trifolium hybridum var. elegans, Trifolium pratense, Trifolium repens, Medicago 
lupulina, Medicago sativa, Melilotus alba, Melilotus officinalis, Acer saccharinum, 
Aesculus glabra, Impatiens pallida, Parthenocissus quinquefolia, Viola sororia, 
Osmorhiza longistylis, Torilis japonica, Conium maculatum, Pastanaca sativa, 
Teucrium occidentale var. boreale, Nepeta Cataria, Glechoma hederacea, Prunella 
vulgaris, Laminum purpureum, Leonurus cardiaca, Solanum carolinense, Ver- 

388 



Plant Taxonomy 389 

bascum Blattaria, Verbascum Thapsus, Campsis radicans, Plantago lanceolata, 
Plantago virginica, Dipsacus sylvestris, Specularia perfoliate^ Ambrosia trifida, 
Ambrosia artemisiifolia var. elatior, Anthemis Cotula, Chrysanthemum Leucan- 
themum var. pinnatifidum, Cirsium arvense, Cirsium discolor, Tragopogon por- 
rifolius. 

Shelby County: Juglans nigra, Carya ovata, Quercus bicolor, Melilotus alba, 
Fraxinus pennsylvanica var. subintegerrima, Ampelamus albidus, Solanum 
carolinense, Linaria vulgaris, Chelone glabra var. linifolia, Cirsium arvense. 

Literature Cited 

1. Deam, C. C. 1940. Flora of Indiana. Indiana Department of Conservation, In- 
dianapolis. 1236 p. 

2. Fernald, M. L. 1950. Gray's Manual of Botany. American Book Co., New 
York. 1632 p. 

3. Keller, C. 1979. Quantitative techniques for the determination of 
phytogeographic areas. Ph.D. dissertation. The University of Notre Dame. 
338 p. 



Vegetation Survey of Hillside Seeps at Turkey Run State Park 

John E. Ebinger, Eastern Illinois University, 

Charleston, Illinois 61920 

and 

John A. Bacone, Indiana Department of Natural Resources, 

Indianapolis, Indiana 46204 



Introduction 

Hillside seeps are small areas, usually less than one-half acre in size, with 
saturated soil caused by seepage water flowing to the surface in a diffuse flow (8). 
They are found most commonly along the lower slopes of glacial moraines and river 
terraces where bands of sand and gravel carry the water to outlet areas forming a 
distinct seepline. The plant community found in these areas commonly contain 
plant species that are rarely encountered in central Indiana. The reason for this 
rare plant community is that the seep water is slightly alkaline and fairly cool (6). 
This unique community has not been studied in detail in central Indiana, though 
similar areas have been examined in central Illinois (3, 6, 7). The present study was 
undertaken to determine if the floristic composition and the abundance of the taxa 
of the Indiana hillside seeps is similar to that found in the seeps of central Illinois. 

Methods 

Each of the seeps was examined three times during the growing season of 
1979, and five times during the growing season of 1980. During each trip the abun- 
dance of each species encountered was recorded. The abundance of a taxon was 
determined by finding the mean distance after a series of measurements were 
made between individuals of the same species (1). The species abundance class was 
then determined by using the scale listed below. In this scale the letter "L" is used 
before the abundance class designation to indicate local abundance. This letter 
means that the species was found in one small, localized part of the seep, and its 
abundance in that area is indicated by the subsequent letters 

DISTANCE 



SYMBOL 


MEANING 


VA 


Very Abundant 


A 


Abundant 


VC 


Very Common 


C 


Common 


VF 


Very Frequent 


F 


Frequent 


F- 


Less Frequent 


FF 


Fairly Frequent 


FO 


Fairly Occasional 





Occasional 



0-3 inches apart 
3-6 inches apart 
6-12 inches apart 
1-1 V2 feet apart 
lVz-2 feet apart 
2-3 feet apart 
3-6 feet apart 
6-12 feet apart 
12-30 feet apart 
30-50 feet apart 



Voucher specimens of all species encountered are deposited in the E. L. 
Stover Herbarium of Eastern Illinois University (EIU), while the nomenclature 
used follows Mohlenbrock (4). 

Description and Location of Seeps 

Both of the seeps studied are located at the eastern edge of Turkey Run State 
Park, Parke County, Indiana, just to the south of the covered bridge (NW l U , S W V4 , 
Sect. 26, T17N, R7W). 



Plant Taxonomy 391 

Area 1: Located on a north-facing hillside to the west of trail one. It is about 37 m 
wide and 40 m long, and has a small stream running through it. Some larger in- 
dividuals and many seedlings of Juniperus virginiana L. are scattered throughout 
the seep along with a few small trees of Fraxinus nigra Marsh, which are found in 
the lower part of the seep. 

Area 2: Located on a north-facing hillside about 300 m to the east of the Lusk Earth 
Fill. It is 82 m long and 26 m wide, and has a small stream forming its northern 
boundary. A few scattered individuals of Salix discolor Muhl. occur in the seep. 

Results and Discussion 

A total of 67 species of herbaceous plants were found in the two seeps. Of 
these taxa, 35 were found in Area 1, while 44 were found in Area 2, with only 12 
species being common to both areas. All of these taxa, along with their abundance 
classes are listed in Tables 1 and 2. Most of the species listed are fairly common 



Table 1. Abundance classes for the plants found in the hillside seep along 

trail one, Turkey Run State Park, Parke County, Indiana (Area 1). For an 

explanation of the abundance class symbols see the Methods section of this 

paper. 

SPECIES ABUNDANCE 

A grostis alba L. C 

Bidens aristosa L. VC 

Carex blanda Dewey. FO 

Carex hystricina Muhl. 

Carex laevivaginata (K § ukenth.) Mack. 

Carex lasiocarpa Ehrh. 

Carex leptalea Wahlenb. VC 

Carex sterilis Willd. A 

Cassia marilandica L. 

Chara brittonii T. F. A. LA 

Clemetis virginiana L. 

Cyperus flavescens L. 

Equisetum arvense L. C 

Eupatorium perfoliatum L. 

Fragaria virginiana Duchesne. 

Gentiana andrewsii Griseb. LF- 

Gerardia tenuifolia Vahl. F - 

Juncus brachycephalus (Engelm.) Bush. C 

Lycopus americanus Muhl. FF 

Lysimachia quadriflora Sims. C 

Mentha X piperita L. 

Oxypolis rigidior (L.) Coulter & Rose. C 

Panicum lanuginosum Ell. FO 

Parnassia glauca Raf. C 

Pedicularis lanceolata Michx. 

Rosa Carolina L. FO 
Rudbeckia fulgida Ait. var. sullivantii (Boynt. & Beadle) 

Cronq. F- 

Scirpus americanus Pers. C 

Scripus pendulus Muhl. 

Selaginella apoda (L.) Fern. LVA 

Senecio pauperculus Michx. 

Solidago patula Muhl. 

Solidago riddellii Frank. F - 

Spiranthes lucida (H. H. Eaton) Ames. LF- 

Viola pratincola Greene. 

Zizia aurea (L.) Koch. 



392 



Indiana Academy of Science 



Table 2. Abundance classes for the plants found in the hillside seep east of 
the Lusk Earth Fill, Turkey Run State Park, Parke County, Indiana (Area 2). 
For an explanation of the abundance class symbols see the Methods section of 

this paper. 



SPECIES 



CAREX 
ZONE 



Acorus calamus L. 
Apios americana Medic. 
Asclepias incarnata L. 
Aster puniceus L. 
Caltha palustris L. 
Cardamine bulbosa (Schreb.) 

BSP. 
Carex hystricina Muhl. 
Carex laevivaginata (K§vkenth.) 

Mack. 
Carex leptalea Wahl. 
Carex shortiana Dewey. 
Carex sterilis Willd. 
Carex torta Boott. 
Chelone glabra L. 
Cirsium altissimum (L.) Spreng. 
Cirsium muticum Michx. 
Cryptotaenia canadensis (L.) 

DC. 
Cuscuta gronovii Willd. 
Eleocharis erythropoda Steud. 
Eupatorium maculatum L. 
Eupatorium perfoliatum L. 
Eupatorium rugosum Houtt. 
Glyceria striata (Lam.) Hitch- 
cock. 
Impatiens biflora Walt. 
Juncus brachycephalus 

(Engelm.) Buch. 
Juncus interior Wieg. 
Leersia oryzoides (L.) Swartz. 
Leersia virginica Willd. 
Liparis loeselii (L.) Rich. 
Lobelia siphilitica L. 
Lycopus virginicus L. 
Lysimachia quadriflora Sims. 
Nasturtium officinale R. Br. 
Oxypolis rigidior (L.) Coulter 

& Rose. 
Rudbeckia fulgida Ait. var. 

sullivantii (Boynt. & 

Beadle) Cronq. 
Rumex obtusifolius L. 
Scirpus atrovirens Willd. 
Scirpus validus Vahl. 
Selaginella apoda (L.) Fern. 
Senecio pauperculus Michx. 
Smilacina stellata (L.) Desf. 
Solidago patula Muhl. 
Symplocarpus foetidus (L.) 

Nutt. 
Thelypteris palustris Schott. 
Vernonia gigantea (Walt.) Trel. 



C 

A 



FO 





VA 
FO 
LC 



FO 




F- 



LVA 





F- 
LVA 
VF 

FF 


FO 



IMPATIENS 


ACORUS 


ZONE 


ZONE 


LF- 


A 





F 


- 


FO 


F- 


F- 


F 


F- 



FF 

FO 



C 
VA 



LVA 




LA 

F 



F- 

LA 






FO 



VC 



FO 



Plant Taxonomy 393 

throughout most of Indiana and Illinois, and would be expected in this type of 
habitat. Eight of these taxa, however, are rarely encountered in west-central In- 
diana (2) and adjacent Illinois (5). For the most part, these taxa are relict species 
that have northern affinities. Included in this group are Carex lasiocarpa, Carex 
leptalea, Carex sterilis, Cirsium muticum, Juncus brachycephalus, Liparis loeselii, 
Parnassia glauca, and Spiranthes lucida. 

The vegetation of the seep along trail one at Turkey Run State Park (Area 1) 
is fairly uniform and distinct vegetation zones are not present. Of the 35 species 
found here (Table 1) nearly half are listed as being fairly occasional or occasional in 
abundance. Of the more abundant species in the seep, members of the Cyperaceae 
dominate, with the most abundant taxon in the seep being Carex sterilis. Other im- 
portant taxa of this family are Carex leptalea and Scirpus americanus. Other taxa 
commonly encountered are Agrostis alba, Bidens aristosa, Equisteum arvense, 
Juncus brachycephalus, Lysimachia quadriflora, Oxypolis rigidior, and Parnassia 
glauca. One interesting species found is the alga, Chara brittonii. This rather rare 
taxon is found in small pools and on moist soil near the upper edge of the seep. 

The flora of Area 2 is much more diverse than that of Area 1 (Table 2). Three 
distinct vegetation zones exist in this seep as a result of differences in shading, 
moisture, and soil conditions. These are the Carex zone, the Impatiens zone, and 
the Acorus zone. The Carex zone, which occurs in most of the northern and central 
parts of the seep is extremely moist, has a thick organic soil, and is rarely shaded. 
It is the most diverse part of the seep, with 28 taxa occurring here. The dominant 
plants of this zone are As ter puniceus, Cardamine bulbosa, Carex leptalea, Carex 
sterilis, Eleocharis erythropoda, Rudbeckia fulgida, and Senecio pauperculus 
(Table 2). The Impatiens zone is also very moist, has a thick organic soil, and is 
heavily shaded. It occurs all along the southern part of the seep, as well as in fairly 
extensive parts of the east and west edges of the seep. Of the 25 taxa found here, 
most are shade tolerant. The dominant species of this zone are Cardamine bulbosa, 
Impatiens biflora, and Symplocarpus foetidus. The Acorus zone, is the smallest 
zone in the seep, and occurs in a slightly raised area in the southeastern part. This 
zone, which is rarely shaded, is slightly drier than the other zones, and has a thin- 
ner soil. Only 13 taxa are found here with Acorus calamus and Impatiens biflora be- 
ing dominant. 

The flora of the two seeps is not very similar. Of the 67 herbaceous taxa en- 
countered, only 12 were found in both areas. For the most part, these floristic dif- 
ferences are caused by soil conditions. In Area 1 the soil is very thin, has a high con- 
centration of clay, and has very little organic material. Also, at the upper part of 
the seep the clay parent material is exposed. Area 2, in contrast, occurs on very 
loose gravel and sand, and has a relatively thick, highly organic soil. 

The floristic composition of Area 2 is similar to that found in many of the seeps 
of east-central Illinois (3). Most of the dominant species are the same, as are some of 
the relict and less common species. Of the taxa found in Area 2, about 70 percent 
are found in the Illinois seeps. The differences are probably a reflection of chance 
dissemination, the amount of disturbances in and around the seeps, and some dif- 
ferences in habitat. Area 1, in contrast, differs from most of the seeps studied in 
east-central Illinois (3). It is similar, however, to one mentioned by Ebinger (3) that 
was found at the base of Windfall Prairie in Vermilion County, Illinois. Both seeps 
have a similar floristic composition, and the soil is similar. 

Acknowledgments 

The authors would like to thank the Indiana Department of Natural Resources 



394 Indiana Academy of Science 

for permission to study the seeps, and to Dr. Fay Kenoyer Daily for the identifica- 
tion of Chara brittonii. 

Literature Cited 

1. Acocks, J. 1953. Veld types of South Africa. Bot. Survey Mem. No. 28, Dept. 
Agric, Div. Bot. Pretoria, Union of South Africa. 

2. Deam, C. C. 1940. Flora of Indiana. Wm. B. Burford Printing Co., Indianapolis. 

3. Ebinger, J. E. 1978. Vascular flora of hillside seeps in east-central Illinois. 
Trans. 111. St. Acad. Sci. 71:109-114. 

4. Mohlenbrock, R. H. 1975. Guide to the vascular flora of Illinois. Southern Il- 
linois University Press, Carbondale and Edwardsville. 

5. Mohlenbrock, R. H. and D. M. Ladd. 1978. Distribution of Illinois vascular 
plants. Southern Illinois University Press, Carbondale and Edwardsville. 

6. Parker, H. and J. E. Ebinger. 1971. Ecological study of a hillside marsh in 
east-central Illinois. Trans. 111. St. Acad. Sci. 64:362-369. 

7. Phipps, R. and J. Speer. 1958. A hillside marsh in east-central Illinois. Trans. 
111. St. Acad. Sci. 51:37-42. 

8. White, J. and M. H. Madany. 1978. Appendix 30. Classification of natural com- 
munities in Illinois, in White, J. natural areas inventory. Technical report. 



Additions to the Flora of Gibson County, Indiana 

William E. McClain 

Natural Heritage Section, Illinois Department of Conservation, 

Springfield, Illinois 62706 

A floristic study was conducted to determine the vascular plant populations of 
northwestern Gibson County, Indiana. This investigation resulted in the collection 
of 35 vascular plant species not previously reported for Gibson County by Deam 
(1940) or subsequent updates to the flora of Gibson County (Deam et al 1940, 1944, 
1947, 1949, 1951; Humbles 1965, 1970; Kays and Humbles 1974). Including the 
species reported here, the vascular flora recorded for Gibson County totals 618 
species: 18 ferns or fern allies, 1 gymnosperm, 166 monocots, and 433 dicots. 

The habitats that were investigated in this study consisted of lowland forest, 
roadsides, fallow fields, sloughs and floodplain forest adjacent to the Wabash 
River. Because the study area is well within the broad, level floodplain of the 
Wabash River, a considerable amount of the land is subjected to annual spring 
flooding. 

Of the 35 species of vascular plants reported here as new to Gibson County, 
none of them represent new additions to the flora of Indiana. Two seldom collected 
plants in Indiana, Cyperus acuminatus T. & G. and Scirpus smithii A. Gray, were 
collected along the shores of a soil borrow area. 

In the following list, the name of each species is followed by a brief note con- 
cerning its habitat and the author's collection numbers. The nomenclature follows 
Fernald (1950). All specimens were deposited in the Deam Herbarium of Indiana 

University. 

Acknowledgment 

Special appreciation is extended to Dr. John Ebinger for the verification and 
identification of specimens and for review of the manuscript. 

Ferns and Fern Allies 
Equisetaceae 

Equisetum hyemale L. Roadside; 2194. 

Salvinaceae 

Azolla caroliniana Willd. Pond; 2517. 

Monocotyledoneae 

Cyperaceae 

Carex lacustris Willd. Roadside; 2389. 

Cyperus acuminatus T. & G. Artificial pond; 2075. 

Scirpus smithii A. Gray Artificial pond; 2074. 

Graminae 

Bromus tectorum L. Fallow Field; 2384. 
Dactylus glomerata L. Roadside; 2392. 
Festuca elatior L. Roadside; 2219. 
Hordeum pusillum Nutt. Roadside; 2381. 



396 Indiana Academy of Science 

Paspalum pubiflorum Rupr. Roadside; 2220. 
Poa chapmaniana Scribn. Roadside; 2386. 
Setaria faberii Herrm. Roadside; 2221. 
Setaria glauca (L.) Beauv. Roadside; 2199. 
Uniola latifolia Michx. Moist Woods; 2226. 

Naidaceae 

Naias guadalupensis (Spreng) Magnus. Artificial pond; 2077. 
Naias minor All. Artificial pond; 2078. 

Dicotyledoneae 
Annonaceae 

Asimina triloba (L.) Dunal. Moist Forest; 2288. 

C ary ophy llaceae 

Silene antirrhina L. Roadside; 2387. 

Compositae 

Aster ontarionis Wieg. Roadside; 2174. 

Aster simplex Willd. Roadside; 2258. 

Bidens aristosa (Michx.) Britt. Fallow Field; 2255. 

Erechitites hieracifolia (L.) Raf. Fallow Field; 2014. 

Cruciferae 

Lepidium virginicum L. Roadside; 2380. 

Dioscoraceae 

Dioscorea villosa L. Moist Forest; 2234. 

Leguminosae 

Lespedeza stipulacea Maxim. Roadside; 2209. 
Medicago lupulina L. Roadside; 2374. 
Robinia psuedoacacia L. Roadside; 2368. 
Trifolium hybridum L. Roadside; 2383. 
Trifolium pratense L. Roadside; 2372. 
Vicia villosa Roth Roadside; 2396. 

Oxalidaceae 

Oxalis dillenii Jacq. Roadside; 2437. 

Phytolaccaceae 

Phytolacca americana L. Roadside: 2184. 

Rosaceae 

Rosa multiflora Thunb. Roadside; 2395. 

Salicaceae 

Salix interior L. Shore of pond; 2388. 

Umbelliferae 

Zizia aurea (L.) Koch Wabash River Floodplain; 2365. 



397 Plant Taxonomy 

Literature Cited 

1. Deam, C. C. 1940. Flora of Indiana. Indiana Department of Conservation, Divi- 
sion of Forestry. Indianapolis. 1236 pp. 

2. Deam. C. C, R. C. Friesner. R. Kriebel and T. G. Yuncker. 1940. Indiana 
Plant Distribution Records. Proc. Indiana Acad. Sci. 50:72-78. 

3. Deam, C. C, R. Kriebel. T. G. Yuncker and R. C. Friesner. 1944. Indiana 
Plant Distribution Records V. Proc. Indiana. Acad. Sci. 54:91-99. 

4. Deam. C. C, T. G. Yuncker and R. C. Friesner. 1947. Indiana Plant Distribu- 
tion Records, VIII. Proc. Indiana Acad, of Sci. 57:81-86. 

5. Deam. C. C, T. G. Yuncker and R. C. Friesner. 1949. Indiana Plant Distribu- 
tion Records, X. Proc. Indiana Acad. Sci. 59:48-52. 

6. Deam. C. C, T. G. Yuncker and R. C. Friesner. 1951. Indiana Plant Distribu- 
tion Records, XII. Proc. Indiana Acad. Sci. 61:72-77. 

7. Fernald. M. L. 1950. Gray's Manual of Botany, eighth edition, American Book 
Company, New York, New York, 1632 pp. 

8. Humbles. J. 1965. Indiana Plant Distribution Records XIX. Proc. Indiana 
Acad. Sci. 75:231-232. 

9. Humbles. J. 1970. Indiana Plant Distribution Records XXI. Proc. Indiana 
Acad. Sci. 80:413. 

10. Kays B. and J. Humbles. 1974. Indiana Plant Distribution Records XXII. Proc. 
Indiana Acad. Sci. 84:428. 



Additions to the Flora of Vermillion County, Indiana 

William E. McClain 

Natural Heritage Section, Illinois Department of Conservation 

Springfield, Illinois 62706 

During 1977 and 1978, the author conducted a study of the vascular plant 
populations in Vermillion County, Indiana. This study resulted in the collection of 
96 vascular plant species not previously reported for Vermillion County by Deam 
(1940) or subsequent updates to the flora of Vermillion County (Deam et al 1945, 
1948; 1950; Buser 1953). Including the species reported here, the vascular flora 
recorded for the county totals 616 species; 11 ferns, 127 monocots, and 478 dicots. 

The habitats included in this study included upland and lowland forests, road- 
sides, marshes and fallow fields. None of the 96 vascular plant species reported 
here as new to Vermillion County represent new additions to the flora of Indiana. 

In the following list of plants, each species is followed by a brief note concern- 
ing its habitat and the author's collection numbers. The nomenclature follows Fer- 
nald (1950). Voucher specimens of all species were deposited in the Deam Her- 
barium of Indiana University. 

Acknowledgment 

A special appreciation is extended to Dr. John Ebinger for the identification 
and verification of specimens and for reviewing the manuscript. 

Monocotyledoneae 
Commelinaceae 

Tradescantia subaspera Ker. Upland Forest: 2057. 

Cyperaceae 

Carex blanda Dewey. Upland Forest; 2285. 
Carex conoidea Schk. Upland Forest; 2277. 
Carex rosea Schk. Upland Forest; 2289. 
Carex stipata Muhl. Marsh; 2295. 

Graminae 

Dactylis glomerata L. Roadside; 2354. 
Eleusine indica L. Gaertn. Fallow Field; 2119. 
Elymus villosus Muhl. Upland Woods; 2435. 
Festuca elatior L. Fallow Field; 2136. 
Poa chapmaniana Schrib. Roadside; 2343. 
Poa pratensis L. Fallow Field; 2309. 
Sorghum vulgare Pers. Roadside; 2146. 

Juncaceae 

Juncus dudley Weig. Marsh; 2470. 

Liliaceae 

Belamcanda chiensis (L.) DC Fallow Field; 2166. 
Camassia scilloides (L.) Desf. Upland Forest; 2319. 
Erythronium albidum Nutt. Upland Forest; 2270. 

398 



Plant Taxonomy 

Polygonatum commutatum Schultes. Upland Forest; 2322. 
Smilax hispida Muhl. Wabash River Flood Plain; 2113. 

Orchidaceae 

Triphora trianthorphora (Sw) Rydb. Upland Forest; 2516. 

Typhaceae 

Typha augustifolia L. Marsh; 2502. 
Typha latifolia L. Marsh; 2503. 

Dicotyledoneae 
Asclepiadaceae 

Asclepias hirtella L. Roadside; 2451. 

Boraginaceae 

Mertensia virginica (L.) Pers. Upland Forest; 2274. 

Caryophyllaceae 

Holosteum umbellatum L. Roadside; 2318. 
Stellaria media (L.) Vill. Roadside; 2300. 

Celastraceae 

Celastrus scandens L. Fallow Field; 2167. 

C he napod iace ae 

Chenopodium album L. Wabash River Floodplain; 2110. 

Commelinaceae 

Tradescantia subaspera Ker. Upland Forest; 2057. 

Compositae 

Arctium minus (Hill) Bernh. Roadside; 2412. 
Aster oblongifolius Nutt. Marsh; 2023. 
Bidens coronata (L.) Britt. Marsh; 2029. 
Cirsium vulgare (Savi) Tenore. Road; 2165. 
Eupatorium serotinum Michx. Roadside; 2049. 
Matricaria matricarioides (Less) Porter. Roadside; 2314. 
Prenanthes altissima L. Upland Forest; 2097. 
Solidago ulmifolia Muhl. Upland Forest; 2099. 
Taraxacum officinale Wiggers. Roadside; 2307. 
Verbesina altemifolia (L.) Britt. Marsh; 2087. 
Verbesina helianthoides Michx. Fallow Field; 2058. 
Vernonia altissima Nutt. Fallow Field; 2465. 

Convolvulaceae 

Cuscuta glomerata Choisy. Parasitic on Solidago; 2448. 
Cuscuta gronovii Willd. Parasitic on Bidens; 2499. 

Cruciferae 

Barbarea vulgaris R.Br. Roadside; 2266. 

Descurainia brachycarpa (Richards) O.E. Schulz. Roadside; 2356. 

Nasturtium officinale R.Br. Marsh; 2340. 



400 Indiana Academy of Science 

Cucurbitaceae 

Sicyos angulatus L. Wabash River Floodplain; 2107. 

Euphorbiaceae 

Acalypha virginica L. Roadside; 2064. 
Euphorbia platyphylla L., Roadside; 2120. 
Euphorbia supina Raf. Roadside; 2142. 

Fumariaceae 

Corydalis flavula Raf. DC. Fallow Field; 2265. 
Dicentra cucullaris (L.) Bernh. Upland Forest; 2281. 

G rossulariaceae 

Ribes americanum Mill. Marsh; 2476. 

Ribes missouriense Nutt. Upland Forest; 2287. 

Hydrophyllaceae 

Hydrophyllum macrophyllum Nutt. Upland Forest; 2324. 
Hydrophyllum virginianum L. Upland Forest; 2338. 

Hypericaceae 

Hypericum punctatum Lam. Upland Forest; 2433. 
Hypericum sphaerocarpum Michx. Upland Forest; 2429. 

Labia tae 

Lamium purpureum L. Roadside; 2264. 
Leonurus cardiaca L. Marsh; 2089. 
Lycopus virginicus L. Roadside; 2196. 
Physostegia speciosa Sweet., Fallow Field; 2158. 
Scutellaria lateriflora L. Marsh; 2496. 

Leguminosae 

Amorpha fruticosa L. Fallow Field; 2028. 
Lespedeza intermedia (Wats) Upland Forest; 2427. 
Robinia psuedoacacia L. Roadside; 2147. 

Lobeliaceae 

Lobelia siphilitica L. Marsh; 2481. 

Malvaceae 

Abutilion theophrasti Medic. Roadside; 2130. 

Oxalidaceae 

Oxalis dillenii Jacq. Upland Forest, 2437. 
Oxalis stricta L. Upland Forest; 2437B. 

Penthoraceae 

Penthorum sedioides L. Marsh; 2497. 

Phytolaccaceae 

Phytolacca americana L. Roadside; 2144. 



Plant Taxonomy 401 

Plantaginaceae 

Plantago rugelii R. & S. Fallow Field; 2121. 

Podophyllaceae 

Podophyllum peltatum L. Upland Forest; 2336. 

Polygonaceae 

Polygonum lapthifolium L. Roadside; 2069. 
Rumex altissimus Wood. Fallow Field; 2118. 

Portulacaceae 

Claytonia virginica L. Upland Forest; 2288. 

Ranunculaceae 

Actaea alba (L.) Mill. Upland Forest; 2514. 
Ranunculus septentrionalis Poir. Fallow Field; 2294. 

Rosaceae 

Potentilla monospliensis L. Roadside; 2413. 
Potentilla simplex Michx. Fallow Field; 2347. 
Rosa setigera Michx. Marsh; 2477. 

Rubiaceae 

Galium obtusum Bigel. Upland Forest; 2428. 
Rutaceae 

Pltea trifoliata L. Upland Forest; 2512. 
Zanthoxylum americanum Mill. Upland Forest; 2330. 

Salicaceae 

Salix discolor Muhl. Marsh; 2291. 

Salix interior Rowlee. Fallow Field; 2357. 

Scrophulariaceae 

Penstemon digitalis Nutt. Fallow Field; 2449. 
Solonaceae 

Datura stramonium L. Fallow Field; 2116. 
Solanum carolinense L. Fallow Field; 2062. 
Solanum nigrum L. Fallow Field; 2122. 

Umbelliferae 

Sium sauve Walt. Marsh; 2495. 
Zizia aurea (L.) Koch. Roadside; 2298. 

Urticaceae 

Parietaria Pennsylvania Muhl. Upland Forest; 2089. 
Pilea fontana (Lunell) Rydb. Marsh; 2035. 

Verbenaceae 

Verbena hastata L. Marsh; 2482. 

Violaceae 

Viola rafinesquii Greene. Roadside; 2258. 



402 Indiana Academy of Science 



Literature Cited 

Buser, F. B. 1953. Indiana Plant Distribution Records, XIV. Proc. of the Ind. 
Acad, of Sci. 63:67-72. 

Deam, C. C. 1940. Flora of Indiana. Indiana Department of Conservation, Divi- 
sion of Forestry. Indianapolis. 1236 pp. 

Deam. C. C, R. Kriebel. T. G. Yuncker, and R. C. Friesner. 1945. Indiana 
Plant Distribution Records, VI. Proc. of the Ind. Acad, of Sci., 45:50-64. 
Deam, C. C, T. G. Yuncker and R. C. Friesner. 1948. Indiana Plant Distribu- 
tion Records, IX. Proc. of the Ind. Acad, of Sci., 58:92-96. 
Deam, C. C, T. G. Yuncker and R. C. Friesner. 1949. Indiana Plant Distribu- 
tion Records X., Proc. of the Ind. Acad, of Sci., 59:48-52. 
Deam, C. C, T. G. Yuncker and R. C. Friesner. 1950. Indiana Plant Distribu- 
tion Records, XL Proc. of the Ind. Acad, of Sci., 60:82-90. 
Fernald, M. L. 1950. Gray's Manual of Botany, Eighth Edition. American 
Book Company. New York, New York 1652 pp. 



SCIENCE EDUCATION 

Chairman: William G. Wert 
Department of Life Sciences, Indiana State University, Terre Haute, Indiana 47809 

Chairman-Elect: Charles Gehring 
Department of Life Sciences, Indiana State University, Terre Haute, Indiana 47809 

ABSTRACTS 

Wabash Summer Aquatic Biology Program -The First Year. Austin E. Brooks 
and William N. Doemel, Department of Biology, Wabash College, Crawfordsville, 

Indiana 47933. The Wabash Summer Aquatic Biology Program provides 

undergraduates a unique opportunity to learn about aquatic microbial ecology and 
camping — wilderness techniques. For each of four summers, students will visit a 
different aquatic ecosystem. This past summer the focus was the extreme en- 
vironments of Yellowstone National Park. In subsequent summers visits will be 
made to the Wisconsin lakes and the Boundary Waters (Mn), the Northeast 
seacoast, and the Southeast seacoast. A 34 foot avion trailer serves as a laboratory. 
Equipped with an epifluorescense and 8 phase microscopes, a liquid scintillation 
counter, spectrophotometers, pH meters and other limnological supplies, this 
laboratory enables students to study microorganisms in their natural habitats and 
to isolate them in pure culture. The course consists of a number of orientation 
laboratories in which students learn microbial techniques. The primary emphasis, 
however, is placed on the development of projects by each pair of students. The 
students sleep in tents and are responsible for preparing all of their meals. 
Although consuming a considerable amount of time each day, through these ac- 
tivities the students learn much about themselves and about cooperation with 
others. Also many informal discussions highlight the significant scientific ex- 
periences of the day. We believe that the concept of a well-equipped mobile 
laboratory and tents — a traveling field course — offers an exciting alternative to 
the traditional field station. 

Botanical and Zoological Prints in the Collections of the Hugh Thomas Miller Rare 
Book Room, Irwin Library, Butler University. Richard A. Davis, University 

Librarian, Butler University, Indianapolis, Indiana 46208. A short history of 

the development of biological illustration which begins with the "Hortus 
Sanitatus" of 1491, and includes examples from such monuments of natural history 
as Fuchs "Historia Stirpium" (1542), Gesner's "Historia Animalium" (1551), 
William Curtis' "Flora Londinensis" (1777), and the frontispiece "Portrait of Lin- 
naeus in Lapland Dress" from Thornton's "Temple of Flora" (1799), as exemplified 
by the collection of original botantical and zoological prints housed in the Hugh 
Thomas Miller Rare Book Room of the Irwin Library. 

This collection of some 80 original graphic works is. in two parts: The Jeanette 
Siron Pelton collection of Botanical prints; and a parallel collection of zoological 
prints. 

The collections were formed during the years 1970-73 with the encouragement 
of Dr. John F. Pelton, Chairman of the Department of Botany at Butler, and a grant 
for library enrichment from the Sears-Roebuck Foundation. 

A Simple Quantitative Method for Teaching Phylogenetic Relationships Among 

403 



404 Indiana Academy of Science 

Animal Phyla to College Freshman. James D. Haddock, Department of Biological 

Sciences, IU-PU at Fort Wayne, Fort Wayne, Indiana 46805. A technique is 

described to enable students in introductory zoology to become better acquainted 
with major phyla of animals and their relationships to one another. In the 
laboratory, students first are asked to determine to what phylum an organism 
belongs by using a traditional taxonomic key. After having correctly identified the 
organism, students then are asked to assign a point value to each phylum based on 
an accompanying chart listing a variety of characteristics. Each attribute, such as 
body symmetry, for example, will have a number of alternatives — each having a 
point value. Each phylum thus receives a total score based on the sums of the ag- 
gregate characteristics. By looking at point values and totals, students can easily 
spot evolutionary trends and identify "simple" and "complex" phyla. Also by obser- 
ving clusters of numbers, even non-science majors can easily achieve a quick, 
overall understanding of phylogenetic relationships among major groups. 

STEP: The First Year: John A. Ricketts, Department of Chemistry, and Paul B. 
Kissinger, Department of Physics, DePauw University, Greencastle, Indiana 
46135. STEP, an acronym for Science Training Enrichment Program, is an at- 
tempt to strengthen science and mathematics education in the secondary school. 
The premise of STEP, which is supported by a three-year grant from the Interna- 
tional Business Machines Corporation, is to create a viable, academic, high school- 
college interface across which science education can be promoted. It is anticipated 
that STEP might serve as a successful model to strengthen the education in the 
high school for any academic discipline. 

The major components of STEP during the first year were: 

(1) The Planning Session: All aspects of STEP were formulated in coopera- 
tion with science teachers from the participating high schools and DePauw science 
faculty during a two-day summer workshop. 

(2) Saturday Science Seminars: One hundred thirty-five high school students 
(and 18 teachers) received 16 hours of instruction in one of the basic areas, Com- 
puters and Mathematics, Physical Sciences, or Biological Sciences on four Satur- 
days, two each semester. 

(3) "Frontiers of Science" Lecture Series: Four eminent scientist/educators 
visited the campus and presented their views. The series was open to the campus 
and the community and also was advertised to the high schools within commuting 
distance of DePauw University. 

(4) Computer Science Module for High School Teachers: Seventeen teachers 
received 16 hours (four successive Saturdays) instruction in programming skills 
and the use of the computer. 

(5) Consultants Service Program: This component provides for visiting lec- 
turers to the high school and also offers the opportunity to the high school to use 
the science facilities at DePauw University. 

(6) Summer Practicum in Science: A two-week total immersion in one of the 
following areas, Physics, Chemistry and Computers, Topics in Mathematics, or In- 
troduction to the Biological and Health Sciences, attracted 48 students from six 
states. 

Formal assessment of the impact of the program will be made at the end of the 
second year. A random sampling of student participants from the two years and 
their science teachers will be invited to campus in order to evaluate the real 
strengths and weaknesses of the program. 



Science Education 405 

Summary Report of Science Educators' Survey of Science Methods Curriculum. 

Stanley S. Shimer, Science Teaching Center, Indiana State University, Terre 

Haute, Indiana. 47809. The purposes for this study were: 1) to identify the 

science educators' consensus regarding the content for the selected topics within 
the elementary school teacher preparation curriculum and 2) to examine the 
science educators' opinions concerning the content that should be included in an 
elementary school teacher education program. The science educators who were 
surveyed in this study (N = 44) were selected from recognized leaders in the field 
throughout the nation. 

The instrument used in this study was the Science Educators Opinion Survey 
(Appendix A) which consisted of two parts. The questionnaire was sent to seventy- 
two (72) science educators located in forty-five (45) states. The mailing consisted of 
a cover letter, the Science Educators Information questionnaire, the Science 
Education Opinion Survey, and a self-addressed, stamped envelope. Three weeks 
after the original mailing a follow-up reminder postcard was sent to all non- 
respondents. 

A panel of forty-four (44) science educators was asked to indicate the best con- 
text in which each of the forty-six (46) curricular topics should be presented. The 
context options provided were: (A) Science Methods; (B) Foundations of Educa- 
tion; (C) Introduction of Education; (D) Educational Psychology; (E) Others and 
(F) Uncertainty. As a first step in analysis, the responses were grouped. The 
designation "A" was used for the responses which indicated the context ot science 
methods only. The group "B" denoted the responses in which science methods plus 
one or more of the other option areas was the recommended context. If the science 
educators' responses recommended that the topic should not be taught in science 
methods, the response was put into group "C". 

The Crosstabulation Procedures and the chi-square test were used to deter- 
mine whether there was agreement regarding the placement of the forty-six (46) 
topics in the curriculum of the pre-service elementary school teacher. 

In conclusion, the science educators did come to an agreement of what should 
be taught in science elementary methods class. The author has made some specific 
recommendations for future science methods teachers. If done again, it also should 
try to identify the trends in science methods preparation for elementary majors. 

The Necessity and Methods of Teaching Communication Skills for Science Fair 
Participants. Patricia Arnett Zeck, Northwestern High School, 3917 North 

Northwestern Road, Kokomo, Indiana 46901. Written and oral self-evaluation 

and discussion by advanced biology students indicate a direct correlation between 
communication skills and success in science fair competition. An increasing number 
of judges is reading the research paper in its entirety. Oral questioning may last as 
long as twenty minutes per judge with a minimum of 3 judges per student. To help 
the student prepare for a successful experience it is necessary for the science 
teacher to guide more than scientific research of each student. He must also aid in 
the library research of each student. He must review reference citation form, 
mathematical analysis and computation, and graphing. He must stress organization 
and presentation of data. He must mark the written paper for grammar and spell- 
ing. He should demonstrate the rudiments of effective speaking through personal 
appearance, poise, gestures, voice control, and elimination of distracting man- 
nerisms. He should help the student arrange a color coordinated display board with 
topics according to the scientific method in order to facilitate an organized analysis 
and presentation. Following these guidelines will help the exhibitor to have 
greater confidence and success in his discussion of his science project. 



SOIL AND ATMOSPHERIC SCIENCES 



Chairman: Donald P. Franzmeier 
Department of Agronomy, Purdue University, West Lafayette, Indiana 47907 

Chairman-Elect: Robert F. Dale 
Department of Agronomy, Purdue University, West Lafayette, Indiana 47907 

ABSTRACTS 

Clay Mineral Study of Soils from the Savannah River Plant South Carolina. E. A. 

Matney and R. V. Ruhe, Water Resources Research Center and Department of 

Geology, Indiana University, Bloomington, Indiana 47405. Six soil samples 

from the Savannah River Plant, located near Aiken, South Carolina, were studied 
to determine the nature of the clay minerals. This paper concentrates on the iden- 
tification and estimation of clay mineral content. Oriented slides of the < 2um frac- 
tion of the soils were x-rayed before and after heating for 15 minutes at 450°C and 
600°C, and after vaporization for 48 hours with ethylene glycol. Kaolinite (7.2 A) 
and a 14 A clay mineral dominated the suite. The 14 A mineral is problematic in 
that it is nonexpansible and poorly collapsible. Upon heating at 450°C and 600°C, 
and 14 A peak collapses to a plateau from 14 to 10 A. After a selective dissolution 
procedure, it is concluded that the 14 A mineral in these soils is an interlayered 
chlorite-vermiculite. Clay mineral content was estimated using a modification of a 
method proposed by Ruhe and Olson. Increasing proportions of soil clay is added to 
a constant standard composed of equal parts by weight of the < 2um fraction of a 
Georgia kaolinite and a Texas montmorillonite. The mixtures were prepared for 
x-ray powder diffraction analysis and x-rayed after vaporization with ethylene 
glycol. Peak areas were measured and intensity ratios calculated. These ratios 
were plotted against °/o soil clay. Estimation equations were derived and used to 
determine the amount of kaolinite and the 14 A clay mineral in each of the soil 
samples. 

Parent Material — Landscape — Soil Interrelationships in Jefferson County, In- 
diana Allan L. Nickell, United States Department of Agriculture, Soil Conserva- 
tion Service, Madison, Indiana 47250 and Stanley M. Totten, Department of 

Geology, Hanover College, Hanover, Indiana 47243. The soils of Jefferson 

County have formed from widely diverse parent materials ranging from limestone, 
dolomite, and shale bedrock to unconsolidated sand, silt, and clay deposited by 
glaciers, streams, and wind. The relationship between parent materials and soil 
morphology is a direct one, and the presence of a particular soil is a strong indica- 
tion of the type of geologic material in which the soil formed. Because of the 
polygenetic origin, soil scientists need to correlate their mapping with that of 
geologists and geomorpholists in the local survey area. 

During the process of conducting the Jefferson County Soil Survey, many 
joint field studies were made by soil scientists, geologists and geomorpholists for 
the purpose of determining interrelationships among parent materials, landscapes 
and soils. Studies were made by transect, some holes were drilled with a core drill- 
ing machine, and much valuable knowledge was also accumulated and recorded in 
notes during the process of mapping as the soil scientists traversed the land. 

The bedrock exposed in Jefferson County belongs to the Ordovician, Silurian, 

406 



Soil and Atmospheric Sciences 407 

and Devonian systems of the Paleozoic Era. These rocks were deposited as fine 
grained shale and limestone in shallow marine waters from about 450 to 350 million 
years ago. 

Jefferson County was covered by continental ice sheets two or three different 
times in the last 1 million years. These glaciers diverted the drainage of 
southeastern Indiana across the divide at Madison to form the Ohio River, but the 
most significant contribution of the glaciers was the deposition of till and outwash 
which are important soil parent materials. 

Melting of Illinoian ice resulted in deposition of terrace gravels along Big 
Creek in the northwestern part of the county. 

The Wisconsinan ice advance did not reach as far south as Jefferson County, 
but meltwaters of this last ice advance deposited large quantities of sand and 
gravel outwash in the Ohio River Valley. 

Following melting of the last of the ice sheets about 14,000 years ago, rivers 
and streams have modified the landscapes slightly, and have cut new floodplains in- 
to the underlying materials. These modern floodplains contain alluvian deposits of 
clay, silt, sand, gravel, and cobbles. 

The joint studies made by geologists, geomorphologists and soil scientists 
have proven the value of cooperating among these disciplines in collecting and 
analyzing field and lab data. 

The Varying Length of the Growing Season in Indiana. Lawrence A. Schaal, 
Department of Agronomy, Purdue University, West Lafayette, Indiana 

47907. The recent trend of colder winters in Indiana causes speculation that the 

length of the frost-free season is becoming shorter. The answer to this question lies 
in the comparison of early and late years of frost data recorded by over twenty 
climatological stations located in Indiana. Data begin in the 19th century. From 
such data we ascertain the short term trend, the intermediate and the over all 
trend of the growing season length. By definition and early years of observations, 
the period from the day of the last freezing temperature (0°C.) in spring to the first 
day of 0°C. or colder in the fall serves to define the frost-free season or the warm 
season crop growth period. Conflicting deductions of growing season length trend 
come from the varying length of historical data used in coming to conclusions. 



Freeze-Thaw Cycles in Indiana Soils 

Robert F. Dale, Beth C. Reinke, and John E. Wright 
Purdue University, West Lafayette, Indiana 47907 

Introduction 

The freezing and thawing of water in soils have both detrimental and 
beneficial effects. The development of potholes in streets and highways are 
familiar to everyone. Newman (1967) measured the heaving of cylindrical fence 
posts caused by freezing and thawing of soil water. Detrimental agricultural 
effects are the heaving of winter cover crops and increases in the soil erosion 
potential. Beneficial agricultural effects of soil freezing and thawing include 
decreasing soil compaction and breaking down limestone and aggregate fertilizer 
materials for faster availability for plant growth. 

For these and other uses, soil temperature records for Indiana were used to 
determine the probabilities of various numbers of freeze-thaw cycles in northern, 
central and southern portions of the state. 

Data and Procedures 
Published daily soil temperature data (USDC, 1965-1980) for selected depths 
under a bare soil surface were used to compute the number of freeze-thaw cycles 
for three locations in Indiana: Purdue Agronomy Farm, West Lafayette 6 NW; 
Southern Indiana Purdue Agricultural Center, Dubois; and Pinney-Purdue 
Agricultural Center, Wanatah 2 WNW. Soil temperatures at 2.5 cm -the 
shallowest and most active freeze-thaw layer in which temperatures were 
measured -were used at all three locations. In addition, soil temperatures for the 
10- 20- and 50-cm depths were also analyzed for West Lafayette 6 N W, the station 
with the longest record. All locations had slopes of 0°, but the soil types varied: 
Toronto silt loam (Udollic Ochraqualf, fine-silty, mixed, mesic) at West Lafayette 
6NW; Zanesville silt loam (Typic Fragiudalf, fine silty, mixed mesic) at Dubois; and 
both Edwards muck (Limnic Medisaprist, marly, euic, mesic) and Tracy sandy loam 
(Ultic hapludalf, course-loamy, mixed, mesic) at Wanatah 2 WNW. 

The time periods used to tabulate the number of freeze-thaw cycles were not 
identical for all locations, since some data were missing and some were suspect. A 
15-year period (fall 1965 to spring 1980) was used for West Lafayette 6NW, ex- 
cluding the 1966-1967 and 1967-1968 seasons for the 10-cm soil depth. A 13-year 
period (fall 1965 to spring 1978) was used for Dubois, and an 11-year penri <&U 
1967 to spring 1980) was used for Wanatah, deleting the 1974-1975 and 1976-1977 
seasons. 

Soil maximum and minimum temperatures were measured with a Palmer dial- 
type thermometer. The Palmer soil thermometers are calibrated twice a year (spr- 
ing and fall) and have an accuracy of ± 0.5°C. An exposure time of about 10 minutes 
is required for accurate registration of a maximum or minimum temperature. Pro- 
per placement of the soil probe is essential for accurate readings. Errors occa- 
sionally are caused by soil erosion or heaving, which effectively change the depth of 
the thermometer, and soil cracking in dry weather, which may expose part of the 
sensor. 

Freeze-thaw cycles were computed from temperature data in the following 
manner. A thaw, T, was defined as a temperature greater than OC and a freeze, F, 

408 



Soil and Atmospheric Sciences 



409 



was defined as a temperature less than or equal to OC. Each season initially began 
with a T. Each pass from T to F, or F to T was considered a half-cycle, with a full 
cycle being two half cycles. Since the observations were taken at 0800 EST, the soil 
temperature was assumed to progress from the maximum temperature of observa- 
tional day 1 to the minimum temperature of observation day 1 to the maximum 
temperature of observational day 2 and to the minimum temperature of observa- 
tional day 2. 

F-T frequencies were compiled within climatological weeks, March 1-7 being 
week 1 and Feburary 21-27 as week 52. FT cycles for February 28 and 29 were in- 
cluded with seasonal totals. The weekly mean numbers of FT cycles were deter- 
mined by summing the number of cycles within climatological weeks over all years. 
The number of F-T cycles expected at the 20, 50, and 80% probability levels were 
estimated by the relation, n + i where m was the rank of the number of F-T cycles 
in ascending order, and n was the number of years of record. 

Results and Discussion 

The number of freeze-thaw cycles at the 2.5-cm soil depth for each winter for 
the four soil-locations — West Lafayette, Dubois, Wanatah-sand, and Wanatah- 
muck — is shown in Figure 1. 

Generally, West Lafayette had the greatest number of freeze-thaw cycles at 
the 2.5-cm depth per season, X = 44.3 cycles/season and with a maximum of 80 in_ 
the 1974-75 season ^Figure 1). Wanatah had an intermediate number of cycles, X 
sand = 31.1 and X muck = 21.6. Dubois had the lowest average number of 
cycles per season, X = 11.2, and there were no soil temperatures OC or below dur- 
ing the 1973-74 and 1974-75 seasons. The absence of freeze-thaw cycles at the 




65/66 66/67 67/68 68/69 69/70 70/71 71/72 72/73 7V74 74/75 75/76 76/77 77/78 78/79 79/80 

WINTER SEASON 

FIGURE 1. Number of freeze-thaw cycles at 2.5-cm depth under bare soil surface for 
indicated season at Dubois and West Lafayette 6NW, and sand and muck soils at 
Wanatah 2WNW, Indiana. 



410 



Indiana Academy of Science 



Dubois 2.5-cm soil depth in these two seasons was unusual. The records were 
checked to see if possibly the Palmer dial-type thermometers were out of calibra- 
tion. The 2.5-cm temperatures appeared consistent with those at 5 and 10 cm. Ap- 
parently, the soil did not freeze at any depth in the several periods during which air 
temperatures were below OC in those seasons, because of high soil moisture, occa- 
sional snow cover, and upward soil heat flux. Central Indiana tends to have more 
freeze-thaw cycles at 2.5 cm than either northern or southern Indiana where soil 
temperatures tend to remain at or below OC and above OC, respectively. 

The observed differences in average number of soil freeze-thaw cycles are 
caused not only by the geographic location of the station, but also by the natural 
drainage, physical, and chemical composition of the soil at each station, as shown by 
the frequencies for the two locations at Wanatah. The sandy soil had a greater 
mean number of freeze-thaw cycles than did the muck soil. Because muck retains 
more water than sand, the muck soil has a higher conductive capacity than the san- 
dy soil, which decreases the soil temperature fluctuations as well as the number of 
freeze-thaw cycles. 

The variation in the number of freeze-thaw cycles from year-to-year was caus- 
ed by the different weather regimes. The number of freeze-thaw cycles usually 
decreased as the mean winter temperature decreased at West Lafayette. This was 
observed in the 77-78 winter which was unusually cold. The minimum number of 
F-T cycles at the 2.5-cm depth for West Lafayette occurred in the 67-68 season (19) 
and the maximum number occurred in the 74-75 season (80), a relatively mild 
winter. 

At West Lafayette 6NW, the number of freeze-thaw cycles for each winter 
season are shown also for the 10-, 20-, and 50-cm depths in Figure 2. Every season 




80 



70 



60 



50 



40 



30 



20 



65/66 66/67 67/68 68/69 69/70 70/71 71/72 72/73 73/74 74/75 75/76 76/77 77/78 78/79 79/80 

WINTER SEASON 

Figure 2. Number of freeze-thaw cycles at the indicated depths under a bare soil 
surface for indicated season at West Lafayette 6NW, Indiana. 



Soil and Atmospheric Sciences 411 

the number of cycles was considerably greater for the 2.5-cm depth than for any of 
the other depths. For example, in the 1976-1977 season there were a total of 53 
freeze-thaw cycles at the 2.5-cm depth, 17 at 10 cm, 6 at 20 cm, and only 1 cycle at 
the 50-cm depth. The maximum number of cycles at 20 cm (30 in the 1972-1973 
season) was greater than that at 10 cm (24 in 1974-1975). This and the greater 
number of freeze-thaw cycles at 20 cm than at 10 cm in the 1971-1972 and 1972-1973 
winter seasons indicate the OC isotherm was oscillating near the 20-cm depth. The 
mean seasonal number of freeze-thaw cycles at the 10-cm depth was 12.5, at 20 cm 
7.3, and at 50 cm 1.1. 

The total number of days with OC or lower for each season and depth are 
shown for West Lafayette 6NW in Figure 3. The shallower depths had more days 
with OC or below. The greatest number of freeze days (69) at the 50-cm depth oc- 
curred during the cold winter of 1976-1977, but in most winters the freezing level 
remained above the 50-cm depth, and in one season, 1973-1974, above the 20-cm 
depth. The fewer number of freezing days at greater depths results from the up- 
ward soil heat flux and the blanketing effect of the top soil layers and, occasionally, 
snow cover. 

The first and last dates of soil freezing are shown for each season and depth at 
West Lafayette 6 NW in Figure 4. The earliest first freeze at the 2.5-cm depth was 
October 18, 1976, and the latest first freeze December 2, 1968. The earliest last 
freeze at the 2.5-cm depth was March 9, 1966, and the latest last freeze was April 
14, 1973. The period of possible soil freezing decreases with greater soil depths. 

The mean number and 80, 50, and 20 percent probabilities of having the in- 
dicated number or fewer freeze-thaw cycles in each week are shown in Figure 5 for 
the 2.5-, 10-, and 20-cm soil depths at West Lafayette 6NW. There is a distinct 
bimodal pattern with the first maximum occurring in the late fall and early winter 




65/66 66/67 67/68 68/69 69/70 70/71 71/72 72/73 73/74 74/75 75/76 76/77 77/78 78/79 79/80 

WINTER SEASON 

Figure 3. Number of days with soil temperatures OC or lower at the indicated 
depth for each season at West Lafayette 6NW, Indiana. 



412 



Indiana Academy of Science 



2.5 CM DEPTH 
10 CM DEPTH 
20 CM DEPTH 
50 CM DEPTH 




2.5 CM DEPTH 
10 CM DEPTH 
20 CM DEPTH 
50 CM DEPTH 



65/66 67/68 69/70 71/72 73/74 75/76 77/78 79/80 
66/67 68/69 70/71 72/73 74/75 76/77 78/79 

WINTER SEASON 
Figure 4. First and last dates of soil freezing at the indicated depth for each 
season at West Lafayette 6NW, Indiana. 



and the second maximum in the late winter and early spring. For example, at the 
2.5-cm depth there was an 80% probability of having 5 or fewer cycles within each 
of the weeks ending November 14 to Dec. 5, 3 or fewer cycles in each week ending 
Dec. 12 to Feb. 13, and 6 or fewer cycles during the weeks ending Feb. 27 to Apr. 
14. The 2.5-cm weekly means peaked at 2.7 cycles in the week ending Dec. 5, and at 
3.9 cycles in the week ending Feb. 27. The means were less than 1.5 cycles from the 
week ending Dec. 6 to that ending Feb. 13. Note that for the skewed distributions, 
bounded at and 7 cycles in each week, the means were above the 50% pro- 
babilities for all but the week ending Nov. 28. The curves for the 10- and 20-cm 
depths in Figure 5 showed a lower number of FT cycles with increasing depth. The 
highest 10-cm weekly mean occurred in the week ending Mar. 7 (1.7). Other means 
generally were less man 1.0 cycle per week. The 10-cm 80% probability showed 3 
or less cycles occurred for the weeks ending Feb. 27 through Mar. 21, and also for 
the week ending Jan. 9. The frequency of freeze-thaw cycles decreases with in- 



WEST LAFAYETTE 
BARE SOIL 



Soil and Atmospheric Sciences 



PROBABILITY OF 
INDICATED NUMBER 
OR FEWER F-T 
CYCLES IN WEEK 



413 

7 




80% 



MEAN 



50% 



20% 



i 4 

i 

UJ O 

1 

Si 


K) CM DEPTH 




\ 


» L. 




x <^' N A 


jr y 


ll \- 
l l i 


^- 80 % 

\ .^MEAN 
--*^\^^50% 


k? 


Sn 


—j i kj d- — i 



3r 



i3 



80 % 



20 CM DEPTH 



sStl 




MEAN 



17 M SI 7 14 21 28 9 12 19 
OCT NOV DEC 



i 30 6 B 20 27 7 14 21 28 4 II 18 
JAN FEB MAR APR 

WEEK ENDING 



Figure 5. Weekly mean and number of freeze-thaw cycles at 80-, 50-, and 
20-percent probability levels at indicated depth for West Lafayette 6NW, Indiana, 
fall 1965-spring 1980, less 1966-67 and 1967-68 seasons. 



creasing depth. All 20-cm weekly means were less than 1 cycle per week, again a 
result of the insulating effect of top soil and, occasionally, snow cover. 

The weekly mean and the probabilities of the number of freeze-thaw cycles for 
each week at the 2.5-cm soil depth are shown in Figure 6 for Dubois. Two sharp 
peaks are visible for the 80% probability, one for 5 or less cycles per week in the 
week ending Jan. 2, and the other for 4.5 cycles or less in the week ending Feb. 27. 
The mean number of weekly freeze-thaw cycles was less than 1.5 for all weeks at 
the 2.5 cm depth, with the exception of 2 cycles or less in the week ending Jan. 2. 
The minimum number of freeze-thaw cycles occurred the first week of February. 



Indiana Academy of Science 



PROBABILITY OF 
INDICATED NUMBER 
OR FEWER F-T 
CYCLES IN WEEK 




17 24 31 7 14 21 28 5 
OCT NOV 



12 19 26 2 9 16 23 30 6 13 20 27 
DEC JAN FEB 

WEEK ENDING 



21 28 
MAR 



APR 



Figure 6. Weekly mean and number of freeze-thaw cycles at 80-, 50-, and 
20-percent probability levels for the 2.5-cm depth at Dubois, Indiana, fall 
1965-spring 1978. 



This reflects the tendency for the 2.5-cm soil depth usually to be frozen by this 
date. 

The weekly probabilities and F-T cycle means for Wanatah 2WNW are shown 
in Figure 7 for the sandy and muck soils at the 2.5-cm depth. Both the sandy and 
muck soils show distinct bimodal patterns, and the sandy soil generally had a 
higher probability of freeze-thaw cycles. At the muck site the weekly means were 
highest in the spring with 3.2 cycles or less for each of the weeks ending Feb. 27 to 
Apr. 11, and with 1.0 or less cycles in the remaining weeks. There were less freeze- 
thaw cycles per week in November and December than in the spring months at the 
muck soil site. 

Summary 

Freezing and thawing processes in the soil have both beneficial and detrimen- 
tal effects in agriculture. To determine the probability of the number of freeze- 
thaw cycles at selected depths for bare soil, soil temperature records were sum- 
marized for the occurrences of OC at three locations in Indiana,- West Lafayette 
6NW, Dubois Indiana Forage Farm, and Wanatah 2WNW-for the 2.5-cm soil 
depth at all three locations, also for and/the 10-, 20-, and 50-cm depths at West 
Lafayette 6NW. At Wanatah, freeze-thaw cycles were summed for both a muck 
and a sandy soil. West Lafayette, the most centrally located site, had the highest 
seasonal mean number of freeze-thaw cycles at the 2.5-cm soil depth, averaging 
44.3 cycles/season with a maximum of 80 in the 1974-75 winter. Dubois, the 
southern-most site, had the lowest average, 11.2, and there were no occurrences in 
the 1973-74 and 74-75 seasons. At West Lafayette the average first freeze at 2.5 cm 
was Nov. 8 and the average last freeze Apr. 3. The mean number of freeze-thaw 
cycles and the 80, 50, and 20 percent probabilities of numbers of freeze-thaw cycles 
for each week were presented for all sites. The probabilities showed a strong 
bimodal pattern, with peaks in the fall and early spring. The mean and probability 
peaks for freeze-thaw cycles in the spring months (late Feb. Mar. and Apr.) are 
higher than the peaks in the fall (Nov. and Dec). 



WANATAH, 

BARE SAND 
2 5 CM DEPTH 



Soil and Atmospheric Sciences 



PROBABILITY OF 
INDICATED NUMBER 
OR FEWER F-T 
CYCLES IN WEEK 



80 % 



MEAN 




50% 



415 

6 



80% 



MEAN 



50% 



- 2 



<7 24 31 

OCT 



9 16 23 30 6 13 20 27 7 
JAN FEB 

WEEK ENDING 



APR 



Figure 7. Weekly mean and number of freeze-thaw cycles at 80-, 50-, and 
20-percent probability levels for the 2.5-cm depth in sand and muck soils at 
Wanatah 2WNW, Indiana, fall 1967-spring 1980, less 1974-75 and 1976-77 seasons. 



The climatology of soil freeze-thaw cycles, shown here for three stations in 
Indiana, can be used for any number of applications in agriculture and engineering. 

Literature Cited 

1. Newman, James E. 1967. Total soil heaving motions at the Southern Indiana 
Forage Farm during the 1965-66 winter season. Purdue Univ. Agric. Expt. 
Sta. Res. Progress Rept. 297. West lafayette, IN. 4 p. 

2. Ohlrogge, A. J. and Sherry Fulk-Bringman. 1980. A new look at fineness of 
grinding agricultural limestone. Agronomy Abstracts 72:173. 

3.